LOCUS       V032774                 7734 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032774
VERSION     V032774
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2414
                     /label="FOXC1(NM_001453)"
                     /note="FOXC1(NM_001453)"
                     /gene="FOXC1"
     CDS             2421..2486
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2510..3098
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3164..3212
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3414..3842
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3856..4185
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4252..5046
                     /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    5222..5343
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5392..5408)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5416..5432
                     /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(5440..5470)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5485..5506
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5794..6382)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6553..7413)
                     /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(7414..7518)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgca ggcgcgctac tccgtgtcca
      781 gccccaactc cctgggagtg gtgccctacc tcggcggcga gcagagctac taccgcgcgg
      841 cggccgcggc ggccgggggc ggctacaccg ccatgccggc ccccatgagc gtgtactcgc
      901 accctgcgca cgccgagcag tacccgggcg gcatggcccg cgcctacggg ccctacacgc
      961 cgcagccgca gcccaaggac atggtgaagc cgccctatag ctacatcgcg ctcatcacca
     1021 tggccatcca gaacgccccg gacaagaaga tcaccctgaa cggcatctac cagttcatca
     1081 tggaccgctt ccccttctac cgggacaaca agcagggctg gcagaacagc atccgccaca
     1141 acctctcgct caacgagtgc ttcgtcaagg tgccgcgcga cgacaagaag ccgggcaagg
     1201 gcagctactg gacgctggac ccggactcct acaacatgtt cgagaacggc agcttcctgc
     1261 ggcggcggcg gcgcttcaag aagaaggacg cggtgaagga caaggaggag aaggacaggc
     1321 tgcacctcaa ggagccgccc ccgcccggcc gccagccccc gcccgcgccg ccggagcagg
     1381 ccgacggcaa cgcgcccggt ccgcagccgc cgcccgtgcg catccaggac atcaagaccg
     1441 agaacggtac gtgcccctcg ccgccccagc ccctgtcccc ggccgccgcc ctgggcagcg
     1501 gcagcgccgc cgcggtgccc aagatcgaga gccccgacag cagcagcagc agcctgtcca
     1561 gcgggagcag ccccccgggc agcctgccgt cggcgcggcc gctcagcctg gacggtgcgg
     1621 attccgcgcc gccgccgccc gcgccctccg ccccgccgcc gcaccatagc cagggcttca
     1681 gcgtggacaa catcatgacg tcgctgcggg ggtcgccgca gagcgcggcc gcggagctca
     1741 gctccggcct tctggcctcg gcggccgcgt cctcgcgcgc ggggatcgca cccccgctgg
     1801 cgctcggcgc ctactcgccc ggccagagct ccctctacag ctccccctgc agccagacct
     1861 ccagcgcggg cagctcgggc ggcggcggcg gcggcgcggg ggccgcgggg ggcgcgggcg
     1921 gcgccgggac ctaccactgc aacctgcaag ccatgagcct gtacgcggcc ggcgagcgcg
     1981 ggggccactt gcagggcgcg cccgggggcg cgggcggctc ggccgtggac gaccccctgc
     2041 ccgactactc tctgcctccg gtcaccagca gcagctcgtc gtccctgagt cacggcggcg
     2101 gcggcggcgg cggcggggga ggccaggagg ccggccacca ccctgcggcc caccaaggcc
     2161 gcctcacctc gtggtacctg aaccaggcgg gcggagacct gggccacttg gcgagcgcgg
     2221 cggcggcggc ggcggccgca ggctacccgg gccagcagca gaacttccac tcggtgcggg
     2281 agatgttcga gtcacagagg atcggcttga acaactctcc agtgaacggg aatagtagct
     2341 gtcaaatggc cttcccttcc agccagtctc tgtaccgcac gtccggagct ttcgtctacg
     2401 actgtagcaa gtttggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     2461 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     2521 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2581 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2641 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2701 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2761 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2821 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2881 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     2941 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3001 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3061 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3121 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3181 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3241 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3301 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3361 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3421 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3481 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3541 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3601 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3661 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3721 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3781 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3841 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3901 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3961 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4021 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4081 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4141 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4201 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4261 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4321 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4381 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4441 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4501 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4561 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4621 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4681 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4741 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4801 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4861 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4921 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4981 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5041 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5101 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5161 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5221 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5281 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5341 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5401 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5461 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7561 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7621 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7681 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
