LOCUS       V018356                 9888 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018356
VERSION     V018356
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             2827..3519
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             3526..4656
                     /label="Acta2(NM_031004)"
                     /note="Acta2(NM_031004)"
                     /gene="Acta2"
     promoter        4676..5175
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5196..5795
                     /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    5809..6397
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6604..7237
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7366..7382)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7390..7406
                     /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(7414..7444)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7459..7480
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7768..8356)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8527..9387)
                     /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(9388..9492)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9540..9674
                     /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 tcgctagcgc taccggactc agatctcgag
     2821 gccaccatga gcgagctgat caaggagaac atgcacatga agctgtacat ggagggcacc
     2881 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     2941 cagaccatga agatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     3001 gctaccagct tcatgtacgg cagcaaagcc ttcatcaacc acacccaggg catccccgac
     3061 ttctttaagc agtccttccc tgagggcttc acatgggaga gaatcaccac atacgaagac
     3121 gggggcgtgc tgaccgctac ccaggacacc agcttccaga acggctgcat catctacaac
     3181 gtcaagatca acggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacacgc
     3241 ggctgggagg ccaacaccga gatgctgtac cccgctgacg gcggcctgag aggccacagc
     3301 cagatggccc tgaagctcgt gggcgggggc tacctgcact gctccttcaa gaccacatac
     3361 agatccaaga aacccgctaa gaacctcaag atgcccggct tccacttcgt ggaccacaga
     3421 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga gatggctgtg
     3481 gccaagtact gcgacctccc tagcaaactg gggcacagag aattcatgtg tgaagaggaa
     3541 gacagcacag ctctggtgtg tgacaatggc tccgggctct gtaaggcggg ctttgctggt
     3601 gatgatgctc ccagggctgt tttcccatcc atcgtgggac gtcccagaca ccagggagtg
     3661 atggttggaa tgggccaaaa ggacagctat gtgggggacg aagcgcagag caagagaggg
     3721 atcctgaccc tgaagtatcc gatagaacac ggcatcatca ccaactggga cgacatggaa
     3781 aagatctggc accactcctt ctataacgag cttcgcgtgg ccccagagga gcatccgacc
     3841 ttgctaacgg aggcgccgct gaaccctaag gccaaccggg agaaaatgac ccagattatg
     3901 tttgagacct tcaatgtccc tgccatgtat gtggctattc aggctgtgct gtccctctat
     3961 gcttctggac gtacaactgg tattgtgctg gactctggag atggcgtgac tcacaacgtg
     4021 cctatctatg agggctatgc tctgcctcat gccatcatgc gtctggactt ggctggccga
     4081 gatctcaccg actacctcat gaagatcctg actgagcgtg gctattcctt cgtgactact
     4141 gctgagcgtg agatcgtccg tgacatcaag gagaagctgt gctatgtcgc tctggacttt
     4201 gaaaatgaga tggccactgc tgcttcctct tcttccctgg agaagagcta cgaactgcct
     4261 gatgggcagg tgatcaccat cgggaatgaa cgcttccgct gcccggagac cctcttccag
     4321 ccatctttca ttggaatgga gtcggcgggc atccacgaaa ccacctataa cagcatcatg
     4381 aagtgtgata ttgacatcag gaaggacctc tatgctaaca acgtcctctc ggggggcact
     4441 accatgtacc caggcattgc tgacaggatg cagaaggaga tcacagccct cgctcccagc
     4501 accatgaaga tcaagattat tgctcctcca gaacgcaaat attctgtctg gatcggcggc
     4561 tccatcctgg cttctctatc taccttccag cagatgtgga tcagcaaaca ggagtatgac
     4621 gaagctggcc cctccattgt ccaccgcaaa tgcttctaat ctagataatt ctaccgggta
     4681 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     4741 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     4801 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     4861 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     4921 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     4981 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     5041 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     5101 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     5161 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     5221 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     5281 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     5341 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     5401 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     5461 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5521 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5581 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5641 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5701 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5761 gcacctggtg catgacccgc aagcccggtg cctgaccgcg tctggaacaa tcaacctctg
     5821 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5881 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5941 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6001 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6061 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6121 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6181 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     6241 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6301 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6361 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     6421 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     6481 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     6541 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     6601 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     6661 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6721 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6781 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     6841 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     6901 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     6961 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     7021 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     7081 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7141 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7201 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7261 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc
     7321 tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7381 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7441 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7501 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7561 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7621 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7681 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7741 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7801 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7861 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7921 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7981 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     8041 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     8101 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8161 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     8221 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8281 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8341 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8401 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8461 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8521 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8581 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8641 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8701 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8761 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8821 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8881 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8941 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9001 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9061 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9121 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9181 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9241 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9301 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9361 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9481 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca
     9541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9661 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     9721 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     9781 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9841 tttttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt
//
