LOCUS       V032405                 7402 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032405
VERSION     V032405
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      880..889
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             889..912
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             916..2586
                     /label="PTBP1(NM_002819)"
                     /note="PTBP1(NM_002819)"
                     /gene="PTBP1"
     CDS             2608..3126
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNG
                     IKANFKIRHNIE"
                     /label="VN173"
     polyA_signal    3164..3388
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3434..3862
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3876..4205
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4272..4871
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5045..5166
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5215..5231)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5239..5255
                     /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(5263..5293)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5308..5329
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5617..6202)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6373..7233)
                     /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(7234..7338)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag ccaccatgga ctacaaagac
      901 gatgacgaca agcttatgga cggcattgtc ccagatatag ccgttggtac aaagcgggga
      961 tctgacgagc ttttctctac ttgtgtcact aacggaccgt ttatcatgag cagcaactcg
     1021 gcttctgcag caaacggaaa tgacagcaag aagttcaaag gtgacagccg aagtgcaggc
     1081 gtcccctcta gagtgatcca catccggaag ctccccatcg acgtcacgga gggggaagtc
     1141 atctccctgg ggctgccctt tgggaaggtc accaacctcc tgatgctgaa ggggaaaaac
     1201 caggccttca tcgagatgaa cacggaggag gctgccaaca ccatggtgaa ctactacacc
     1261 tcggtgaccc ctgtgctgcg cggccagccc atctacatcc agttctccaa ccacaaggag
     1321 ctgaagaccg acagctctcc caaccaggcg cgggcccagg cggccctgca ggcggtgaac
     1381 tcggtccagt cggggaacct ggccttggct gcctcggcgg cggccgtgga cgcagggatg
     1441 gcgatggccg ggcagagccc cgtgctcagg atcatcgtgg agaacctctt ctaccctgtg
     1501 accctggatg tgctgcacca gattttctcc aagttcggca cagtgttgaa gatcatcacc
     1561 ttcaccaaga acaaccagtt ccaggccctg ctgcagtatg cggaccccgt gagcgcccag
     1621 cacgccaagc tgtcgctgga cgggcagaac atctacaacg cctgctgcac gctgcgcatc
     1681 gacttttcca agctcaccag cctcaacgtc aagtacaaca atgacaagag ccgtgactac
     1741 acacgcccag acctgccttc cggggacagc cagccctcgc tggaccagac catggccgcg
     1801 gccttcggtg cacctggtat aatctcagcc tctccgtatg caggagctgg tttccctccc
     1861 acctttgcca ttcctcaagc tgcaggcctt tccgttccga acgtccacgg cgccctggcc
     1921 cccctggcca tcccctcggc ggcggcggca gctgcggcgg caggtcggat cgccatcccg
     1981 ggcctggcgg gggcaggaaa ttctgtattg ctggtcagca acctcaaccc agagagagtc
     2041 acaccccaaa gcctctttat tcttttcggc gtctacggtg acgtgcagcg cgtgaagatc
     2101 ctgttcaata agaaggagaa cgccctagtg cagatggcgg acggcaacca ggcccagctg
     2161 gccatgagcc acctgaacgg gcacaagctg cacgggaagc ccatccgcat cacgctctcg
     2221 aagcaccaga acgtgcagct gccccgcgag ggccaggagg accagggcct gaccaaggac
     2281 tacggcaact cacccctgca ccgcttcaag aagccgggct ccaagaactt ccagaacata
     2341 ttcccgccct cggccacgct gcacctctcc aacatcccgc cctcagtctc cgaggaggat
     2401 ctcaaggtcc tgttttccag caatgggggc gtcgtcaaag gattcaagtt cttccagaag
     2461 gaccgcaaga tggcactgat ccagatgggc tccgtggagg aggcggtcca ggccctcatt
     2521 gacctgcaca accacgacct cggggagaac caccacctgc gggtctcctt ctccaagtcc
     2581 accatctcta gaagatccat cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2641 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2701 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     2761 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc
     2821 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2881 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2941 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3001 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3061 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     3121 atcgagtaga gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3181 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3241 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3301 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3361 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3421 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3481 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3541 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3601 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3661 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3721 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3781 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3841 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3961 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4021 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4081 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4141 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4201 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4261 gatcgtttcg catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4321 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4381 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4441 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4501 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4561 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4621 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4681 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4741 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4801 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4861 ccggtgcctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4921 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4981 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5041 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5101 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5161 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5221 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5281 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5341 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6121 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     6181 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6241 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6301 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6361 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6421 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6481 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6541 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6601 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6661 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6721 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6781 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6841 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6901 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6961 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7021 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7081 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7141 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7201 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7261 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7321 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7381 agatctcccg atcccctatg gt
//
