LOCUS       V018876                 9372 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018876
VERSION     V018876
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             2807..3523
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Aequorea coerulescens GFP"
                     /transl_table=1
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
                     /label="AcGFP1"
     CDS             3560..4165
                     /label="TWIST1(NM_000474)"
                     /note="TWIST1(NM_000474)"
                     /gene="TWIST1"
     promoter        4207..4706
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4727..5326
                     /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    5340..5928
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6135..6768
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6897..6913)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6921..6937
                     /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(6945..6975)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6990..7011
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7299..7887)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8058..8918)
                     /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(8919..9023)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9071..9205
                     /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 tctaccggtc gccaccatgg tgagcaaggg
     2821 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagatctcg
     3541 agctcaagct tcgaattcga tgatgcagga cgtgtccagc tcgccagtct cgccggccga
     3601 cgacagcctg agcaacagcg aggaagagcc agaccggcag cagccgccga gcggcaagcg
     3661 cgggggacgc aagcggcgca gcagcaggcg cagcgcgggc ggcggcgcgg ggcccggcgg
     3721 agccgcgggt gggggcgtcg gaggcggcga cgagccgggc agcccggccc agggcaagcg
     3781 cggcaagaag tctgcgggct gtggcggcgg cggcggcgcg ggcggcggcg gcggcagcag
     3841 cagcggcggc gggagtccgc agtcttacga ggagctgcag acgcagcggg tcatggccaa
     3901 cgtgcgggag cgccagcgca cccagtcgct gaacgaggcg ttcgccgcgc tgcggaagat
     3961 catccccacg ctgccctcgg acaagctgag caagattcag accctcaagc tggcggccag
     4021 gtacatcgac ttcctctacc aggtcctcca gagcgacgag ctggactcca agatggcaag
     4081 ctgcagctat gtggctcacg agcggctcag ctacgccttc tcggtctgga ggatggaggg
     4141 ggcctggtcc atgtccgcgt cccactaggg atccaccgga tctagataac tgatcataat
     4201 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     4261 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     4321 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     4381 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     4441 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     4501 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     4561 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     4621 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     4681 cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga ccgagtacaa
     4741 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4801 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4861 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4921 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4981 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5041 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5101 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5161 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5221 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5281 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaccgc gtctggaaca
     5341 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5401 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5461 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5521 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5581 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5641 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5701 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     5761 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5821 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5881 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt
     5941 ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     6001 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     6061 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6121 agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     6181 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     6241 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     6301 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     6361 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     6421 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     6481 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     6541 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     6601 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6661 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6721 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6781 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     6841 cttgacattg ctagcgtttt accgtcgacc tctagctaga gcttggcgta atcatggtca
     6901 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6961 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7021 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7081 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7141 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7201 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7261 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7321 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7381 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7441 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7501 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7561 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7621 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7681 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7741 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7801 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7981 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8041 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8101 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8161 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8221 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8281 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8341 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8401 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8461 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8521 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8581 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8641 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8701 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8761 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8821 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8881 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8941 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9001 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9061 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9181 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     9241 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     9301 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     9361 aaacttagta gt
//
