LOCUS       V028754                 6932 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028754
VERSION     V028754
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             941..2422
                     /label="Foxc2(NM_013519)"
                     /note="Foxc2(NM_013519)"
                     /gene="Foxc2"
     CDS             2429..2494
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2532..2756
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2802..3230
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3244..3573
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3640..4434
                     /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    4608..4729
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4778..4794)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4802..4818
                     /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(4826..4856)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4871..4892
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5180..5765)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5936..6796)
                     /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(6797..6901)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcaggcgc gttactcggt
      961 atcggacccc aacgccctgg gagtggtacc ctatttgagt gagcaaaact actaccgggc
     1021 ggccggcagc tacggcggca tggccagccc catgggcgtc tactccggcc acccggagca
     1081 gtacggcgcc ggcatgggcc gctcctacgc gccctaccac caccagcccg cggcgcccaa
     1141 ggacctggtg aagccgccct acagctatat agcgctcatc accatggcga tccagaacgc
     1201 gccagagaag aagatcactc tgaacggcat ctaccagttc atcatggacc gtttcccctt
     1261 ctaccgcgag aacaagcagg gctggcagaa cagcatccgc cacaacctgt cactcaatga
     1321 gtgcttcgtg aaagtgccgc gcgacgacaa gaagccgggc aagggcagct actggacgct
     1381 cgacccggac tcctacaaca tgttcgagaa tggcagcttc ctgcggcggc ggcggcgctt
     1441 caagaagaag gatgtgccca aggacaagga ggagcgggcc cacctcaagg agccgccctc
     1501 gaccacggcc aagggcgctc cgacagggac cccggtagct gacgggccca aggaggccga
     1561 gaagaaagtc gtggttaaga gcgaggcggc gtcccccgcg ctgccggtca tcaccaaggt
     1621 ggagacgctg agccccgagg gagcgctgca ggccagtccg cgcagcgcat cctccacgcc
     1681 cgcaggttcc ccagacggct cgctgccgga gcaccacgcc gcggcgccta acgggctgcc
     1741 cggcttcagc gtggagacca tcatgacgct gcgcacgtcg cctccgggcg gcgatctgag
     1801 cccagcggcc gcgcgcgccg gcctggtggt gccaccgctg gcactgccat acgccgcagc
     1861 gccacccgcc gcttacacgc agccgtgcgc gcagggcctg gaggctgcgg gctccgcggg
     1921 ctaccagtgc agtatgcggg ctatgagtct gtacaccggg gccgagcggc ccgcgcacgt
     1981 gtgcgttccg cccgcgctgg acgaggctct gtcggaccac ccgagcggcc ccggctcccc
     2041 gctcggcgcc ctcaacctcg cagcgggtca ggagggcgcg ttgggggcct cgggtcacca
     2101 ccaccagcat cacggccacc tccacccgca ggcgccaccg cccgccccgc agccccctcc
     2161 cgcgccgcag cccgccaccc aggccacctc ctggtatctg aaccacggcg gggacctgag
     2221 ccacctcccc ggccacacgt ttgcaaccca acagcaaact ttccccaacg tccgggagat
     2281 gttcaactcg caccggctag gactggacaa ctcgtccctc ggggagtccc aggtgagcaa
     2341 tgcgagctgt cagctgccct atcgagctac gccgtccctc taccgccacg cagcccccta
     2401 ctcttacgac tgcaccaaat acctcgagga ctacaaagac catgacggtg attataaaga
     2461 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     2521 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2581 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2641 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2701 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2761 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2821 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2881 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2941 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3001 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3061 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3121 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3181 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3241 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3421 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3481 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3541 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3601 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3661 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3721 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3781 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3841 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3901 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3961 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4021 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4081 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4141 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4201 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4261 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4321 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4381 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4441 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4501 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4561 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4621 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4681 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4741 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4801 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4861 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4921 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4981 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5701 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5761 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5821 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5881 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5941 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6001 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6061 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6121 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6181 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6241 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6301 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6361 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6421 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6481 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6541 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6601 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6661 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6721 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6781 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6841 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6901 gcacatttcc ccgaaaagtg ccacctgacg tc
//
