LOCUS       V042891                 9370 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042891
VERSION     V042891
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..3282
                     /label="NUDT1(NM_002452)"
                     /note="NUDT1(NM_002452)"
                     /gene="NUDT1"
     CDS             3313..4032
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     misc_feature    4041..4613
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4615..5325
                     /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    5339..5927
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6134..6767
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6895..6911)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6919..6935
                     /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(6943..6973)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6988..7009
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7297..7885)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8056..8916)
                     /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(8917..9021)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9069..9203
                     /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 caccatgggc
     2821 gcctccaggc tctataccct ggtgctggtc ctgcagcctc agcgagttct cctgggcatg
     2881 aaaaagcgag gcttcggggc cggccggtgg aatggctttg ggggcaaagt gcaagaagga
     2941 gagaccatcg aggatggggc taggagggag ctgcaggagg agagcggtct gacagtggac
     3001 gccctgcaca aggtgggcca gatcgtgttt gagttcgtgg gcgagcctga gctcatggac
     3061 gtgcatgtct tctgcacaga cagcatccag gggacccccg tggagagcga cgaaatgcgc
     3121 ccatgctggt tccagctgga tcagatcccc ttcaaggaca tgtggcccga cgacagctac
     3181 tggtttccac tcctgcttca gaagaagaaa ttccacgggt acttcaagtt ccagggtcag
     3241 gacaccatcc tggactacac actccgcgag gtggacacgg tcggcggtgg aggcagtggt
     3301 ggaggcggta gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3361 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3421 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3481 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3541 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3601 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3661 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3721 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3781 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3841 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3901 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3961 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4021 ctgtacaagt aaggatcccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     4081 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     4141 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     4201 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     4261 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     4321 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     4381 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     4441 acctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     4501 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc
     4561 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggtg
     4621 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     4681 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4741 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     4801 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     4861 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     4921 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     4981 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     5041 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     5101 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     5161 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     5221 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     5281 gagggccgcc actccaccgg cggcatggac gagctgtaca agtgaacgcg tctggaacaa
     5341 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5401 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5461 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5521 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5581 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5641 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5701 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     5761 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5821 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5881 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc
     5941 tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     6001 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     6061 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6121 gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     6181 tggatctacc acacacaagg ctacttccct gattagcaga actacacacc agggccaggg
     6241 gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gccagataag
     6301 gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg
     6361 atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat
     6421 cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac
     6481 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg
     6541 cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt
     6601 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6661 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6721 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6781 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc
     6841 ttgacattgc tagcgtttac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6901 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7081 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7981 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8041 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8101 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8161 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8221 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8281 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8341 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8401 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8461 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8521 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8581 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8641 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8701 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8761 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8821 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8881 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8941 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9001 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     9061 gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     9121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     9181 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
     9241 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
     9301 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
     9361 acttagtagt
//
