LOCUS       V031669                 9160 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031669
VERSION     V031669
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"
     CDS             2297..3202
                     /label="Sparc(NM_009242)"
                     /note="Sparc(NM_009242)"
                     /gene="Sparc"
     CDS             3209..3274
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3319..3530
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3543..3811
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3840..3849
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3870..4532
                     /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             4602..4655
                     /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             4656..5255
                     /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    5256..5843
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5917..6150
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6222..6343
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6362..6497
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6535..6551)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6559..6575
                     /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(6583..6613)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6628..6649
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6937..7525)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7696..8556)
                     /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(8557..8661)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9135..9151
                     /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 tagcgaattc gccaccatga gggcctggat cttctttctc ctttgcctgg ccgggagggc
     2341 cctggcagcc cctcagcaga ctgaagttgc tgaggagata gtggaggagg aaaccgtggt
     2401 ggaggagaca ggggtacctg tgggtgccaa cccagtccag gtggaaatgg gagaatttga
     2461 ggacggtgca gaggaaacgg tcgaggaggt ggtggctgac aacccctgcc agaaccatca
     2521 ttgcaaacat ggcaaggtgt gtgagctgga cgagagcaac acccccatgt gtgtgtgcca
     2581 ggaccccacc agctgccctg ctcccattgg cgagtttgag aaggtatgca gcaatgacaa
     2641 caagaccttc gactcttcct gccacttctt tgccaccaag tgcaccctgg agggcaccaa
     2701 gaagggccac aagctccacc tggactacat cggaccatgc aaatacatcg ccccctgcct
     2761 ggattccgag ctgactgaat tccctctgcg catgcgtgac tggctcaaaa atgtcctggt
     2821 caccttgtac gagagagatg agggcaacaa cctcctcact gagaagcaga agctgcgtgt
     2881 gaagaagatc catgagaatg agaagcgcct ggaggctgga gaccaccccg tggagctgtt
     2941 ggcccgagac tttgagaaga actacaatat gtacatcttc cctgtccact ggcagtttgg
     3001 ccagctggat cagcacccta ttgatgggta cctgtcccac actgagctgg ccccactgcg
     3061 cgctcccctc atccccatgg aacattgcac cacacgtttc tttgagacct gtgacctaga
     3121 caacgacaag tacattgccc tggaggaatg ggccggctgc tttggcatca aggagcagga
     3181 catcaacaag gatctggtga tcggatccga ctacaaagac catgacggtg attataaaga
     3241 tcatgacatc gactacaagg atgacgatga caagtgagcg gccgcgaagg atctgcgatc
     3301 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     3361 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     3421 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     3481 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc
     3541 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
     3601 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
     3661 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
     3721 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
     3781 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg
     3841 ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg
     3901 gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg
     3961 gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc
     4021 tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga
     4081 accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg
     4141 aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg
     4201 gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga
     4261 tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg
     4321 gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca
     4381 gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt
     4441 tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc
     4501 agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg
     4561 ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc
     4621 taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc
     4681 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4741 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4801 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4861 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4921 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4981 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5041 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5101 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5161 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5221 gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt
     5281 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5341 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5401 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5461 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5521 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     5581 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5641 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     5701 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5761 gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct tcgccctcag acgagtcgga
     5821 tctccctttg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     5881 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     5941 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6001 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6061 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6121 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6181 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6361 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6421 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6481 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6541 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6601 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8701 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8761 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     8821 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     8881 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     8941 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9001 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9061 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9121 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
