LOCUS       V034859                 9864 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034859
VERSION     V034859
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      2809..2818
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2818..2841
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2848..4503
                     /label="Aire(NM_009646)"
                     /note="Aire(NM_009646)"
                     /gene="Aire"
     misc_feature    4535..5107
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5109..5819
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    5833..6421
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6628..7261
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7389..7405)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7413..7429
                     /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(7437..7467)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7482..7503
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7791..8379)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8550..9410)
                     /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(9411..9515)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9563..9697
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggac
     2821 tacaaggacg acgatgacaa gctcgagatg gcaggtgggg atggaatgct acgccgtctg
     2881 ctgaggctgc accgcaccga gatcgcggtg gccatagaca gtgcctttcc gctgctgcat
     2941 gctctagccg accacgacgt ggtccctgag gacaagttcc aggagacgct ccgtctgaag
     3001 gagaaggaag gctgccccca ggccttccac gccctgctgt cctggctcct gacccgggac
     3061 agtggggcca tcctggattt ctggaggatt ctctttaagg actacaatct ggagcggtac
     3121 agccgcctgc atagcatcct ggacggcttc ccaaaagatg tggacctaaa ccagtcccgg
     3181 aaagggagaa agccccttgc tggtcccaag gccgcggtac tgccacccag accccccacc
     3241 aagagaaaag cactggagga gcctcgagcc accccaccag caactctggc ctcaaagagc
     3301 gtctccagcc caggctccca cctgaagact aagcccccta agaagccaga tggcaacttg
     3361 gagtcacagc accttcctct tggaaacgga attcagacca tggcagcttc tgtccagaga
     3421 gctgtgaccg tggcctctgg ggatgttcca ggaacccgag gggccgtgga agggatcctt
     3481 atccagcagg tgtttgagtc aggaagatcc aagaagtgca ttcaggttgg gggagagttt
     3541 tatacaccca acaagttcga agaccccagt ggcaatttga agaacaaggc ccggagtggt
     3601 agcagcctaa agccagtggt ccgagccaag ggagcccagg tcactatacc tggtagagat
     3661 gagcagaaag tgggccagca gtgtggggtt cctccccttc catccctccc cagtgagccc
     3721 caggttaacc agaagaacga ggatgagtgt gccgtgtgcc acgacggagg tgagctcatc
     3781 tgttgtgacg gctgtccccg ggccttccac ctggcttgcc tgtccccacc tctgcaggag
     3841 atccccagtg gcctctggag atgctcctgc tgcctccagg gcagagtcca acagaacctg
     3901 tcccagcctg aggtgtccag gcccccggag ctacctgcag agaccccgat cctcgtggga
     3961 ctgaggtcag cttcagagaa aaccaggggc ccatccaggg agctcaaagc cagctctgat
     4021 gctgctgtca catatgtgaa cctgctggcc ccgcaccctg cagctcctct gctggagcct
     4081 tcagcactgt gccctctact gagtgctggg aatgaggggc ggccaggtcc agcaccaagc
     4141 gcgcgatgca gtgtgtgtgg cgatggcacc gaggtgttgc ggtgtgcaca ctgtgccgct
     4201 gccttccact ggcgctgcca cttcccgacg gccgccgccc ggccggggac caatctccgc
     4261 tgcaaatcct gctctgcaga ctcgactccc acgccaggca caccgggcga agctgtaccc
     4321 acctctgggc cccgtccagc acctgggctt gccaaggtag gggacgactc tgctagtcac
     4381 gaccctgttc tacataggga cgacctggag tccctcctca atgagcactc atttgacggc
     4441 atcctgcagt gggccatcca gagcatgtca cgcccgctgg ccgagacacc acccttctct
     4501 tcctgaacta gttctagagc ggccgcggat cccgcccctc tccctccccc ccccctaacg
     4561 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     4621 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     4681 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     4741 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     4801 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     4861 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     4921 gagtcaaatg gctcacctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     4981 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     5041 ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     5101 atgataatat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     5161 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     5221 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     5281 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     5341 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     5401 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     5461 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     5521 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     5581 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     5641 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     5701 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     5761 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtgaa
     5821 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5881 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5941 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6001 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6061 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6121 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6181 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6241 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6301 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6361 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6421 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
     6481 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     6541 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     6601 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     6661 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     6721 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6781 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     6841 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6901 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     6961 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     7021 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     7081 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7141 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7201 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7261 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7321 gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg
     7381 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7441 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7501 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7681 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7741 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7981 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8221 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8341 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8401 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8461 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8521 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8641 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8701 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     9421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     9481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9541 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
     9601 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     9661 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
     9721 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
     9781 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
     9841 aatatgtact acaaacttag tagt
//
