LOCUS       V035723                 7778 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035723
VERSION     V035723
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             905..2476
                     /label="Cyp1a1(NM_009992)"
                     /note="Cyp1a1(NM_009992)"
                     /gene="Cyp1a1"
     CDS             2483..2548
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2558..2614
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2627..3346
                     /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"
     polyA_signal    3378..3602
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3648..4076
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4090..4419
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4486..5280
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5454..5575
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5624..5640)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5648..5664
                     /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(5672..5702)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5717..5738
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6026..6611)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6782..7642)
                     /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(7643..7747)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 caccatgcct tccatgtatg gacttccagc cttcgtgtca gccacagagc tgctcctggc
      961 tgtcaccgta ttctgccttg gattctgggt ggtcagagcc acaagaacct gggttcccaa
     1021 aggcctgaag actccaccag ggccctgggg cttgcccttc attggtcaca tgctgactgt
     1081 ggggaagaac ccacatctgt cactgacacg gctgagtcag cagtatgggg acgtgctgca
     1141 gatccgcatc ggctccactc ctgtggtggt gctgagcggc ctgaacacca tcaagcaggc
     1201 cctggtgagg cagggagatg acttcaaggg ccggccagac ctctacagct tcacacttat
     1261 cactaatggc aagagcatga cttttaaccc agactctgga cccgtgtggg ctgcccgccg
     1321 gcgcctggcc cagaatgccc tgaagagctt ctccatagcc tcggacccga cgtcagcatc
     1381 ctcttgctac ttggaggagc acgtgagcaa ggaggctaac tatctcgtca gcaaacttca
     1441 gaaggtgatg gcagaggttg gccactttga cccttacaag tatttggtcg tgtcagtagc
     1501 caatgtcatc tgtgccatat gctttggcca acgttatgac catgatgacc aagagctgct
     1561 cagcatagtc aatctgagca atgagtttgg ggaggttact ggctctggat acccagctga
     1621 cttcattcct gtcctccgtt acctgcctaa ctcttccctg gatgccttca aggacttgaa
     1681 tgataagttc tacagcttca tgaagaagtt aatcaaagag cactacagga catttgagaa
     1741 gggccacatc cgggacatca cagacagcct cattgagcat tgtcaggaca ggaagctgga
     1801 cgagaatgcc aatgtccagc tgtcagatga taaggtcatc acgattgttt tggacctctt
     1861 tggagctggg tttgacacag tcacaactgc tatctcgtgg agcctcatgt acctggtaac
     1921 caaccctagg gtacagagaa agatccagga ggaactagac acagtgattg gcagagatcg
     1981 gcagccccgg ctttctgaca gacctcagct gccctatctg gaggccttca ttctggagac
     2041 cttccggcat tcatccttcg tccccttcac catcccccac agcaccacaa gagatacaag
     2101 tctgaatggc ttctatatcc ccaaggggtg ctgtgtcttt gtgaaccagt ggcaggttaa
     2161 ccatgaccgg gaactgtggg gtgacccaaa cgagttccgg cctgaaaggt ttctcacccc
     2221 cagcggcact ctggacaagc gcttgagtga gaaggtcact ctctttggtt tgggcaagcg
     2281 aaagtgcatc ggagagacca ttggccgatc ggaggtcttt ctcttcctgg ccatcttgct
     2341 gcagcaaata gaatttaagg tgtctccagg ggagaaggtg gatatgactc ctacctatgg
     2401 gctgacttta aaacacgccc gctgtgaaca cttccaagtg cagatgcggt cttctggtcc
     2461 tcagcatctt caggcttcta gagactacaa agaccatgac ggtgattata aagatcatga
     2521 catcgactac aaggatgacg atgacaaggg aagcggagcc actaacttct ccctgttgaa
     2581 acaagcaggg gatgtcgaag agaatcccgg gccaccggtc gccaccatgg tgagcaaggg
     2641 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2701 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2761 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2821 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2881 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2941 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3001 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3061 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3121 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3181 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3241 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3301 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagggc ccgtttaaac
     3361 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3421 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3481 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3541 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3601 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3661 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3721 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3781 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3841 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3901 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3961 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4021 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4081 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4141 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4201 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4261 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4321 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4381 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4441 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4501 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4561 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4621 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4681 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4741 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4801 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4861 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4921 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4981 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5041 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5101 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5161 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5221 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5281 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5341 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5401 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5461 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5521 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5581 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5641 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5701 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5761 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5821 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5881 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5941 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6001 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6061 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6541 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7081 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7381 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7621 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7681 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7741 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
