LOCUS       V026847                 9430 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026847
VERSION     V026847
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      2279..2288
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2288..2359
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     CDS             2366..3544
                     /label="HPD(NM_002150)"
                     /note="HPD(NM_002150)"
                     /gene="HPD"
     promoter        3589..3800
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3813..4081
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4110..4119
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4140..4802
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4872..4925
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4926..5525
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5526..6113
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6187..6420
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6492..6613
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6632..6767
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6805..6821)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6829..6845
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6853..6883)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6898..6919
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7207..7795)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7966..8826)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8827..8931)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9405..9421
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 caccatggac tacaaggatg acgatgacaa ggattacaaa gacgacgatg ataaggacta
     2341 taaggatgat gacgacaaag aattcatgac gacttacagt gacaaagggg caaagcctga
     2401 gagaggccga ttcctccact tccactctgt gaccttctgg gttggcaacg ccaagcaggc
     2461 cacgtcattc tactgcagca agatgggctt tgaacctcta gcctacaggg gcctggagac
     2521 cggttcccgg gaggtggtca gccatgtaat caaacaaggg aagattgtgt ttgtcctctc
     2581 ctcagcgctc aacccctgga acaaagagat gggcgatcac ctggtgaaac acggtgacgg
     2641 agtgaaggac attgcgttcg aggtggaaga ttgtgactac atcgtgcaga aagcacggga
     2701 acggggcgcc aaaatcatgc gggagccctg ggtagagcaa gacaagtttg ggaaggtgaa
     2761 gtttgctgtg ctgcagacgt atggggacac cacacacacc ctggtggaga agatgaacta
     2821 catcggccaa ttcttgcctg gatatgaggc cccagcgttc atggaccccc tacttcctaa
     2881 actgcccaaa tgcagtctgg agatgatcga ccacattgtg ggaaaccagc ctgatcagga
     2941 gatggtgtcc gcctccgaat ggtacctgaa aaacctgcag ttccaccgct tctggtccgt
     3001 ggatgacacg caggtgcaca cggaatatag ctctctgcga tccattgtgg tggccaacta
     3061 tgaagagtcc atcaagatgc ccatcaatga gccagcgcct ggcaagaaga agtcccagat
     3121 ccaggaatat gtggactata acgggggcgc tggggtccag cacatcgctc tcaagaccga
     3181 agacatcatc acagcgattc gccacttgag agagagaggc ctggagttct tatctgttcc
     3241 ctccacgtac tacaaacaac tgcgggagaa gctgaagacg gccaagatca aggtgaagga
     3301 gaacattgat gccctggagg agctgaaaat cctggtggac tacgacgaga aaggctacct
     3361 cctgcagatc ttcaccaaac cggtgcagga ccggcccacg ctcttcctgg aagtcatcca
     3421 gcgccacaac caccagggtt ttggagccgg caacttcaac tcactgttca aggctttcga
     3481 ggaggagcag aacctgcggg gtaacctcac caacatggag accaatgggg tggtgcccgg
     3541 catgtaagcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
     3601 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
     3661 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     3721 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     3781 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
     3841 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
     3901 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     3961 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     4021 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
     4081 ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc
     4141 ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg
     4201 tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca
     4261 aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc
     4321 acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg
     4381 gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca
     4441 gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc
     4501 ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc
     4561 tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg
     4621 acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc
     4681 ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca
     4741 gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg
     4801 ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca
     4861 ccggatctcg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     4921 gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4981 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5041 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5101 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5161 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5221 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5281 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5341 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5401 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5461 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5521 cctgaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5581 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5641 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5701 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5761 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5821 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5881 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5941 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6001 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6061 gtctccgcct tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac
     6121 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6181 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg
     6241 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6301 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6361 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6421 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6481 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6541 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6601 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     6661 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6721 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     6781 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7621 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8941 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     9001 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     9061 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     9121 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     9181 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     9241 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9301 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9361 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9421 tgccaagctg
//
