LOCUS       V043027                 9853 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043027
VERSION     V043027
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        131..334
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             404..2140
                     /label="Gpc3(NM_016697)"
                     /note="Gpc3(NM_016697)"
                     /gene="Gpc3"
     CDS             2186..2896
                     /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    2936..3053
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        3102..3612
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             3634..4233
                     /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"
     LTR             4361..4594
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    4666..4787
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      4827..4962
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(4983..5001)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(5011..5027)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      5169..5624
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5650..5754
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5755..6615
                     /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"
     rep_origin      6786..7374
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7662..7683
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7698..7728
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7736..7752
                     /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"
     primer_bind     7760..7776
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        7797..7815
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        7843..8069
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             8070..8250
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    8297..8422
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    8915..9148
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             9333..9377
                     /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"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg actcagatct cgagctcaag cttcgaagcc accatggccg ggaccgtgcg
      421 caccgcgtgc ttgctggtgg cgatgctgct aggcttgggc tgcctgggac aggcgcagcc
      481 cccgccgcct ccagacgcca cctgtcacca ggtccgttct ttcttccaga gactgcagcc
      541 cggactcaaa tgggttccag aaacccctgt accaggatca gatttgcaag tatgtctccc
      601 caagggccca acatgctgct caagaaagat ggaagaaaaa taccaactaa cagcacggct
      661 gaacatggaa caactgctcc agtctgcgag tatggaactc aagttcttaa ttattcagaa
      721 tgctgcggtt ttccaagagg cctttgaaat tgttgttcgc catgccaaga actacaccaa
      781 cgccatgttc aagaataact accccagcct gactccacaa gcttttgagt ttgtcggtga
      841 atttttcaca gatgtgtctc tctacatctt gggttctgat atcaacgtgg atgatatggt
      901 caatgaattg ttcgacagcc tctttccagt catctacacc cagatgatga acccaggcct
      961 gcctgagtca gtcttagaca tcaacgagtg cctccgagga gcaagacgtg acctgaaagt
     1021 atttggcagt ttccccaagc ttattatgac ccaggtttcc aagtcactgc aagtcactcg
     1081 aatcttcctt caagccctga atctcggaat tgaagtcatc aacactaccg accacctcaa
     1141 gtttagtaag gactgtggcc gtatgctcac ccgaatgtgg tattgctctt actgccaggg
     1201 actgatgatg gttaagcctt gcggtggtta ttgcaatgtg gtcatgcaag gctgtatggc
     1261 tggtgtggtg gagatcgaca agtactggag agaatacatt ctgtctcttg aagagctcgt
     1321 gaatggcatg tacagaatct acgacatgga gaatgtgctg ctcggcctct tttctaccat
     1381 ccatgattcc atccagtatg tgcagaagaa cggaggcaag ctgaccacca ccattggcaa
     1441 gttgtgtgcc cactcccagc aacgccaata tagatctgct tattaccctg aagatctgtt
     1501 tattgacaag aagatattaa aagtcgctca tgtcgaacat gaagaaacct tatccagccg
     1561 aagaagggaa ctgattcaga aactgaagtc tttcatcaac ttctatagcg ctttgccggg
     1621 ctacatctgc agccatagcc ccgtggccga aaatgatacc ctgtgctgga acggacaaga
     1681 acttgtggag agatacagcc agaaggcggc aaggaacggg atgaagaatc agtttaacct
     1741 ccatgagctg aaaatgaagg gccctgagcc ggtggttagc cagatcattg acaaactgaa
     1801 gcacattaac cagctcctga gaaccatgtc tgtgcccaag ggtaaagttc tggataaaag
     1861 cctggatgaa gaaggacttg aaagtggaga ctgcggtgat gatgaagatg aatgcattgg
     1921 aagctctggt gacgggatgg tgaaagtgaa gaatcaactg cgcttccttg cagaactggc
     1981 ctatgatctg gatgtggacg atgctccggg gaacaagcag catggaaatc agaaggacaa
     2041 cgagatcacc acctctcaca gcgtggggaa catgccgtcc ccactgaaga tcctcatcag
     2101 tgtggccatc tatgtggcgt gctttttttt cctggtgcac ggtggaggcg gatcaggagg
     2161 cggtggatct gcaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat
     2221 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     2281 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     2341 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     2401 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     2461 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     2521 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     2581 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     2641 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     2701 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     2761 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     2821 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     2881 cgagctgtac aagtgaattc tcgacctcga gacaaatggc agtattcatc cacaatttta
     2941 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     3001 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     3061 attacaggga cagcagagat ccactttggc cgcggctcga gggggttggg gttgcgcctt
     3121 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     3181 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     3241 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     3301 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     3361 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     3421 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     3481 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     3541 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     3601 ctctctcccc agggggatcc accggagctt accatgaccg agtacaagcc cacggtgcgc
     3661 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3721 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     3781 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3841 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     3901 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     3961 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4021 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4081 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4141 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     4201 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     4261 accgaaagga gcgcacgacc ccatgcatcg gtacctttaa gaccaatgac ttacaaggca
     4321 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     4381 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     4441 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4501 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4561 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     4621 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     4681 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4741 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     4801 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4861 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4921 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     4981 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5041 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5101 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5161 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5221 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5281 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5341 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5401 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5461 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5521 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5581 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     5641 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5701 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5761 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5821 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5881 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5941 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6001 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6061 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6121 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6181 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6241 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6301 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6361 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6421 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6481 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6541 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6601 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6661 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6721 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6781 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6901 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7081 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7141 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7201 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7261 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7321 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7381 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7441 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7501 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7561 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7621 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7681 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7741 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     7801 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     7861 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     7921 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     7981 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     8041 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     8101 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     8161 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     8221 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     8281 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     8341 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     8401 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     8461 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     8521 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     8581 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     8641 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     8701 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     8761 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     8821 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     8881 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     8941 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     9001 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     9061 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     9121 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     9181 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     9241 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     9301 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     9361 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     9421 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     9481 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     9541 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     9601 gatccattcg attagtgaac ggatctcgac ggtatcgatc acgagactag cctcgagcgg
     9661 ccgccccctt caccgagggc ctatttccca tgattccttc atatttgcat atacgataca
     9721 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     9781 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     9841 aaatggacta tca
//
