LOCUS       V024211                 9440 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024211
VERSION     V024211
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    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3483
                     /label="PSMC6(NM_002806)"
                     /note="PSMC6(NM_002806)"
                     /gene="PSMC6"
     CDS             3490..3555
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3599..3810
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3823..4091
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4120..4129
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4150..4812
                     /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             4882..4935
                     /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             4936..5535
                     /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    5536..6123
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6197..6430
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6502..6623
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6642..6777
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6815..6831)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6839..6855
                     /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(6863..6893)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6908..6929
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7217..7805)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7976..8836)
                     /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(8837..8941)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9415..9431
                     /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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg cggaccctag agataaggcg
     2341 cttcaggact accgcaagaa gttgcttgaa cacaaggaga tcgacggccg tcttaaggag
     2401 ttaagggaac aattaaaaga acttaccaag cagtatgaaa agtctgaaaa tgatctgaag
     2461 gccctacaga gtgttgggca gatcgtgggt gaagtgctta aacagttaac tgaagaaaaa
     2521 ttcattgtta aagctaccaa tggaccaaga tatgttgtgg gttgtcgtcg acagcttgac
     2581 aaaagtaagc tgaagccagg aacaagagtt gctttggata tgactacact aactatcatg
     2641 agatatttgc cgagagaggt ggatccactg gtttataaca tgtctcatga ggaccctggg
     2701 aatgtttctt attctgagat tggagggcta tcagaacaga tccgggaatt aagagaggtg
     2761 atagaattac ctcttacaaa cccagagtta tttcagcgtg taggaataat acctccaaaa
     2821 ggctgtttgt tatatggacc accaggtacg ggaaaaacac tcttggcacg agccgttgct
     2881 agccagctgg actgcaattt cttaaaggtt gtatctagtt ctattgtaga caagtacatt
     2941 ggtgaaagtg ctcgtttgat cagagaaatg tttaattatg ctagagatca tcaaccatgc
     3001 atcattttta tggatgaaat agatgctatt ggtggtcgtc ggttttctga gggtacttca
     3061 gctgacagag agattcagag aacgttaatg gagttactga atcaaatgga tggatttgat
     3121 actctgcata gagttaaaat gatcatggct acaaacagac cagatacact ggatcctgct
     3181 ttgctgcgtc caggaagatt agatagaaaa atacatattg atttgccaaa tgaacaagca
     3241 agattagaca tactgaaaat ccatgcaggt cccattacaa agcatggtga aatagattat
     3301 gaagcaattg tgaagctttc ggatggcttt aatggagcag atctgagaaa tgtttgtact
     3361 gaagcaggta tgttcgcaat tcgtgctgat catgattttg tagtacagga agacttcatg
     3421 aaagcagtca gaaaagtggc tgattctaag aagctggagt ctaaattgga ctacaaacct
     3481 gtgggatccg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     3541 gatgacgatg acaagtagcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg
     3601 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     3661 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     3721 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     3781 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     3841 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     3901 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     3961 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     4021 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     4081 cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag
     4141 agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag
     4201 ttcgagctgg tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg
     4261 aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac
     4321 ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc
     4381 aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac
     4441 gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc
     4501 accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc
     4561 gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc
     4621 agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc
     4681 gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag
     4741 gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc
     4801 atcgccttcg ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga
     4861 cccggctcca ccggatctcg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag
     4921 gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4981 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     5041 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     5101 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5161 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5221 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5281 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5341 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5401 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5461 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5521 aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     5581 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5641 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5701 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5761 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5821 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5881 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     5941 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6001 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6061 cctcttccgc gtctccgcct tcgccctcag acgagtcgga tctccctttg gccgcctccc
     6121 cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6181 aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt
     6241 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6301 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6361 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6421 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     6481 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     6541 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6601 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     6661 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6721 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6781 acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt
     6841 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6901 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6961 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7021 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7081 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7141 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7621 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7681 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7741 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7801 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7861 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7921 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8041 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8161 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8401 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8641 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8821 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8881 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8941 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     9001 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     9061 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     9121 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     9181 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     9241 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     9301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     9361 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     9421 cgacggccag tgccaagctg
//
