LOCUS       V036193                 9077 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036193
VERSION     V036193
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..2353
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2366..3793
                     /label="Nsun6(NM_001165941)"
                     /note="Nsun6(NM_001165941)"
                     /gene="Nsun6"
     promoter        3880..4091
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4104..4372
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4401..4410
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4431..5093
                     /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"
     misc_feature    5172..5760
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5834..6067
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6139..6260
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6279..6414
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6452..6468)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6476..6492
                     /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(6500..6530)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6545..6566
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6854..7442)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7613..8473)
                     /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(8474..8578)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9052..9068
                     /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 attctagcgc
     2281 caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaaaga
     2341 cgatgacgac aagcgcgggt ctagaatgta tgtatttcca aagatatctt tgagacctga
     2401 agttgaaaac tatctcaaag agagcttttt gaacgaggag gctgtgtctg cttcaagcag
     2461 acaagaagca gaaaggaaat ttgagactct gctgcttcgc ctgtcacacc ctccgtcgat
     2521 gacaactgtc agagtgaata cgcatttagg ctcagttcag catgtcagag gtctgctgct
     2581 tgaagagctc cagaagcagt ttggtgaatc cagcattcct gttgttcagc atccagccct
     2641 cccggatgtc ttgctcatcc ccatgactgg accaaggaag aatatagaaa gacaacaggg
     2701 tgaagtgatt gttggcgcac agtgtggcaa tgcagtgtta agaggggccc atgtctatgt
     2761 tccaggaatt gtgtcagctt caaaatttat gaaagctgga gatgttattt ccgtgtattc
     2821 tgatattaac ggcaaatgta agaaaggagc caaagaattt gatggaacta aagtatttct
     2881 cggaaacgga atttctgaac tgagccgcaa agatatcttc aatgggttac ctgatctgaa
     2941 aggtataggc ataagaatga cagaacccat atatctcagt ccctcgtttg acaatgttct
     3001 acccagttat atatttttac agaatttacc atctacagtt gtagctcatg ttcttgatcc
     3061 tcaacctgga gagaagattc tagatatgtg tgcagcccct gggggcaaaa ctacacatac
     3121 tgcagcactg atgcaggaca agggagaagt aatagcattg gacaaaatat tgacaaaagt
     3181 gaacaaactg aagcagaatg cttcattatt agggcttcat tctatccggg cattttgctt
     3241 cgatgcaaca aaggcactta agcttgatac taccgatggc atagaagggg ggcctccatt
     3301 cttaccagaa tcttttgacc gaattatcct tgatgcaccc tgcagtggaa tgggacagag
     3361 gccaaatatg gcttgtactt ggactttaaa ggaagtgaca tcctatcagc cactacagcg
     3421 aaaactcctt catgtggcag ttcaactcct gaagccaggg ggagtgctgg tctacagcac
     3481 atgcacaata accttggcag aaaatgaaga gcaagttgcc tgggccctcc gaacattccc
     3541 ctgccttcag cttcaacccc aggaaccaca gattggagga gaagggatgg tgggagctgg
     3601 cctgacattg gagcaattga agcagctgca gcgatttgat ccgtctgttg taccactgca
     3661 gaacatggac actgattctc ttggagaggc cagaagggaa gacatgattt ggttggccaa
     3721 caaagactgt ataggcttct ttattgcaaa gtttctaaaa tgtcaaagca caaaagcgaa
     3781 ggtttctcag aaatgagaat tcgaatttaa atcgggatcc gcggccgcct aattaattaa
     3841 ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     3901 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     3961 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     4021 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     4081 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     4141 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     4201 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     4261 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     4321 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     4381 gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg cccgccatgg
     4441 agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg gtgggcggcg
     4501 gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc aaaggcgccc
     4561 tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac cacttcggca
     4621 cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc ggctacacca
     4681 acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc agctaccgct
     4741 acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc cccgaggaca
     4801 gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac ctgcacccca
     4861 tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc gacggcggct
     4921 actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac cccagcatcc
     4981 tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg
     5041 agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc gccagatccc
     5101 gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc accggatctc
     5161 gctaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5221 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5281 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5341 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5401 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5461 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5521 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5581 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5641 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5701 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5761 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5821 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     5881 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5941 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6001 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6061 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6121 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6181 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6241 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     6301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6361 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     6421 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6481 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6541 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6601 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6661 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6721 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6781 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6841 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6901 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6961 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7021 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7081 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7141 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7201 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7261 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7321 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7381 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7441 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7501 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7561 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7621 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7681 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7741 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7801 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7861 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7921 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7981 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8041 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8101 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8161 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8221 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8281 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8341 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8401 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8461 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8581 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8641 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8701 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8761 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8821 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8881 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8941 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     9001 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     9061 cggccagtgc caagctg
//
