LOCUS       V031951                 7631 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031951
VERSION     V031951
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..3121
                     /label="CTCF(NM_006565)"
                     /note="CTCF(NM_006565)"
                     /gene="CTCF"
     CDS             3128..3193
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3231..3455
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3501..3929
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3943..4272
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4339..5133
                     /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    5307..5428
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5477..5493)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5501..5517
                     /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(5525..5555)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5570..5591
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5879..6464)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6635..7495)
                     /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(7496..7600)
                     /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 atggaaggtg atgcagtcga
      961 agccattgtg gaggagtccg aaacttttat taaaggaaag gagagaaaga cttaccagag
     1021 acgccgggaa gggggccagg aagaagatgc ctgccactta ccccagaacc agacggatgg
     1081 gggtgaggtg gtccaggatg tcaacagcag tgtacagatg gtgatgatgg aacagctgga
     1141 ccccaccctt cttcagatga agactgaagt aatggagggc acagtggctc cagaagcaga
     1201 ggctgctgtg gacgataccc agattataac tttacaggtt gtaaatatgg aggaacagcc
     1261 cataaacata ggagaacttc agcttgttca agtacctgtt cctgtgactg tacctgttgc
     1321 taccacttca gtagaagaac ttcagggggc ttatgaaaat gaagtgtcta aagagggcct
     1381 tgcggaaagt gaacccatga tatgccacac cctacctttg cctgaagggt ttcaggtggt
     1441 taaagtgggg gccaatggag aggtggagac actagaacaa ggggaacttc caccccagga
     1501 agatcctagt tggcaaaaag acccagacta tcagccacca gccaaaaaaa caaagaaaac
     1561 caaaaagagc aaactgcgtt atacagagga gggcaaagat gtagatgtgt ctgtctacga
     1621 ttttgaggaa gaacagcagg agggtctgct atcagaggtt aatgcagaga aagtggttgg
     1681 taatatgaag cctccaaagc caacaaaaat taaaaagaaa ggtgtaaaga agacattcca
     1741 gtgtgagctt tgcagttaca cgtgtccacg gcgttcaaat ttggatcgtc acatgaaaag
     1801 ccacactgat gagagaccac acaagtgcca tctctgtggc agggcattca gaacagtcac
     1861 cctcctgagg aatcacctta acacacacac aggtactcgt cctcacaagt gcccagactg
     1921 cgacatggcc tttgtgacca gtggagaatt ggttcggcat cgtcgttaca aacacaccca
     1981 cgagaagcca ttcaagtgtt ccatgtgcga ttacgccagt gtagaagtca gcaaattaaa
     2041 acgtcacatt cgctctcata ctggagagcg tccgtttcag tgcagtttgt gcagttatgc
     2101 cagcagggac acatacaagc tgaaaaggca catgagaacc cattcagggg aaaagcctta
     2161 tgaatgttat atttgtcatg ctcggtttac ccaaagtggt accatgaaga tgcacatttt
     2221 acagaagcac acagaaaatg tggccaaatt tcactgtccc cactgtgaca cagtcatagc
     2281 ccgaaaaagt gatttgggtg tccacttgcg aaagcagcat tcctatattg agcaaggcaa
     2341 gaaatgccgt tactgtgatg ctgtgtttca tgagcgctat gccctcatcc agcatcagaa
     2401 gtcacacaag aatgagaagc gctttaagtg tgaccagtgt gattacgctt gtagacagga
     2461 gaggcacatg atcatgcaca agcgcaccca caccggggag aagccttacg cctgcagcca
     2521 ctgcgataag accttccgcc agaagcagct tctcgacatg cacttcaagc gctatcacga
     2581 ccccaacttc gtccctgcgg cttttgtctg ttctaagtgt gggaaaacat ttacacgtcg
     2641 gaataccatg gcaagacatg ctgataattg tgctggccca gatggcgtag agggggaaaa
     2701 tggaggagaa acgaagaaga gtaaacgtgg aagaaaaaga aagatgcgct ctaagaaaga
     2761 agattcctct gacagtgaaa atgctgaacc agatctggac gacaatgagg atgaggagga
     2821 gcctgccgta gaaattgaac ctgagccaga gcctcagcct gtgaccccag ccccaccacc
     2881 cgccaagaag cggagaggac gaccccctgg cagaaccaac cagcccaaac agaaccagcc
     2941 aacagctatc attcaggttg aagaccagaa tacaggtgca attgagaaca ttatagttga
     3001 agtaaaaaaa gagccagatg ctgagcccgc agagggagag gaagaggagg cccagccagc
     3061 tgccacagat gcccccaacg gagacctcac gcccgagatg atcctcagca tgatggaccg
     3121 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3181 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3241 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3301 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3361 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3421 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3481 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3541 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3601 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3661 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3721 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3781 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3841 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3901 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4021 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4081 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4141 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4201 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4261 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4321 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4381 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4441 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4501 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4561 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4621 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4681 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4741 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4801 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4861 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4921 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4981 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5041 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5101 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5161 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5221 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5281 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5341 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5401 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5461 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5521 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     6421 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6481 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6541 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6601 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6661 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6721 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6781 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6841 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6901 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6961 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7021 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7081 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7141 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7201 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7261 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7321 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7381 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7441 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7501 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7561 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7621 cacctgacgt c
//
