LOCUS       V021398                 9084 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021398
VERSION     V021398
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"
     CDS             2815..3735
                     /label="NMI(NM_004688)"
                     /note="NMI(NM_004688)"
                     /gene="NMI"
     misc_feature    3755..4327
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4329..5039
                     /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    5053..5641
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5848..6481
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6609..6625)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6633..6649
                     /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(6657..6687)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6702..6723
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7011..7599)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7770..8630)
                     /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(8631..8735)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8783..8917
                     /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 caccatggaa
     2821 gctgataaag atgacacaca acaaattctt aaggagcatt cgccagatga atttataaaa
     2881 gatgaacaaa ataagggact aattgatgaa attacaaaga aaaatattca actaaagaag
     2941 gagatccaaa agcttgaaac ggagttacaa gaggctacca aagaattcca gattaaagag
     3001 gatattcctg aaacaaagat gaaattctta tcagttgaaa ctcctgagaa tgacagccag
     3061 ttgtcaaata tctcctgttc gtttcaagtg agctcgaaag ttccttatga gatacaaaaa
     3121 ggacaagcac ttatcacctt tgaaaaagaa gaagttgctc aaaatgtggt aagcatgagt
     3181 aaacatcatg tacagataaa agatgtaaat ctggaggtta cggccaagcc agttccatta
     3241 aattcaggag tcagattcca ggtttatgta gaagtttcta aaatgaaaat caatgttact
     3301 gaaattcctg acacattgcg tgaagatcaa atgagagaca aactagagct gagcttttca
     3361 aagtcccgaa atggaggcgg agaggtggac cgcgtggact atgacagaca gtccgggagt
     3421 gcagtcatca cgtttgtgga gattggagtg gctgacaaga ttttgaaaaa gaaagaatac
     3481 cctctttata taaatcaaac ctgccataga gttactgttt ctccatacac agaaatacac
     3541 ttgaaaaagt atcagatatt ttcaggaaca tctaagagga cagtgcttct gacaggaatg
     3601 gaaggcattc aaatggatga agaaattgtg gaggatttaa ttaacattca ctttcaacgg
     3661 gcaaagaatg gaggtggaga agtagatgtg gtcaagtgtt ctctaggtca acctcacata
     3721 gcatactttg aagaataggc ggccgcggat cccgcccctc tccctccccc ccccctaacg
     3781 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3841 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3901 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3961 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     4021 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     4081 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     4141 gagtcaaatg gctcacctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     4201 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     4261 ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     4321 atgataatat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     4381 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     4441 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     4501 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     4561 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     4621 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     4681 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     4741 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     4801 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     4861 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     4921 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     4981 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtgaa
     5041 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5101 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5161 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5221 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5281 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5341 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5401 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     5461 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5521 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5581 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5641 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
     5701 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     5761 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     5821 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     5881 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     5941 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6001 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     6061 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6121 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     6181 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     6241 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     6301 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6361 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6421 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6481 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6541 gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg
     6601 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6661 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6721 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6781 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6841 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6901 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6961 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7021 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7081 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7141 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7201 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7261 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7321 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7381 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7441 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7501 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7561 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7621 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7681 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7741 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7801 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7861 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7921 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7981 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8041 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8101 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8161 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8221 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8281 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8341 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8401 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8461 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8521 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8581 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8641 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8701 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8761 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
     8821 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     8881 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
     8941 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
     9001 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
     9061 aatatgtact acaaacttag tagt
//
