LOCUS       V035206                 7410 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035206
VERSION     V035206
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..2090
                     /label="CTBP2(NM_001329)"
                     /note="CTBP2(NM_001329)"
                     /gene="CTBP2"
     CDS             2097..2162
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2186..2774
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2840..2888
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3090..3518
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3532..3861
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3928..4722
                     /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    4898..5019
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5068..5084)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5092..5108
                     /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(5116..5146)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5161..5182
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5470..6058)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6229..7089)
                     /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(7090..7194)
                     /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 ccaccatggc ccttgtggat aagcacaaag
      781 tcaagagaca gcgattggac agaatttgtg aaggtatccg cccccagatc atgaacggcc
      841 ccctgcaccc ccgccccctg gtggcgctgc tggacggccg cgactgcact gtggagatgc
      901 ccatcctgaa ggacctggcc actgtggcct tctgtgacgc gcagtcgacg caggaaatcc
      961 acgagaaggt tctaaacgaa gccgtgggcg ccatgatgta ccacaccatc accctcacca
     1021 gggaggacct ggagaagttc aaggccctga gagtgatcgt gcggataggc agtggctatg
     1081 acaacgtgga catcaaggct gccggcgagc tcggaattgc cgtgtgcaac atcccgtctg
     1141 cagccgtgga agagacagcg gactctacca tctgccacat cctcaacctg taccggagga
     1201 acacgtggct gtaccaggca ctgcgggaag gcacgcgggt tcagagcgtg gagcagatcc
     1261 gcgaggtggc ctcgggagcg gcccgcatcc gtggggagac gctgggcctc attggctttg
     1321 gtcgcacggg gcaggcggtt gcagttcgag ccaaggcctt tggattcagc gtcatatttt
     1381 atgaccccta cttgcaggat gggatcgagc ggtccctggg cgtgcagagg gtctacaccc
     1441 tgcaggattt gctgtatcag agcgactgcg tctccttgca ctgcaatctc aacgaacata
     1501 accaccacct catcaatgac tttaccataa agcagatgag gcagggagca ttccttgtga
     1561 acgcagcccg tggcggcctg gtggacgaga aagccttagc acaagccctc aaggagggca
     1621 ggatacgagg ggcagccctc gacgtgcatg agtcagagcc cttcagcttt gctcagggtc
     1681 cgttgaaaga tgccccgaat ctcatctgca ctcctcacac tgcctggtac agtgagcagg
     1741 cgtcactgga gatgagggag gcagctgcca ccgagatccg ccgagccatc acaggtcgca
     1801 tcccagaaag cttaagaaat tgtgtgaaca aggaattctt tgtcacatca gcgccttggt
     1861 cagtaataga ccagcaagca attcatcctg agctcaatgg tgccacatac agatatccgc
     1921 caggcatcgt gggtgtggct ccaggaggac ttcctgcagc catggaaggg atcatccctg
     1981 gaggcatccc agtgactcac aacctcccga cagtggcaca tccttcccaa gcgccctctc
     2041 ccaaccagcc cacaaaacac ggggacaatc gagagcaccc caacgagcaa ggatccgact
     2101 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     2161 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     2221 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2281 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2341 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2401 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2461 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2521 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2581 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2641 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2701 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2761 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     2821 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     2881 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     2941 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3001 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     3061 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3121 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3181 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3241 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3301 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3361 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3421 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     3481 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3541 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3661 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3721 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3781 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3841 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3901 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3961 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4021 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4081 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4141 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4201 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4261 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4321 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4381 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4441 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4501 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4561 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4621 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4681 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4741 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4861 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5041 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5101 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5161 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5281 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5641 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5881 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7201 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     7261 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7321 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7381 accgacaatt gcatgaagaa tctgcttagg
//
