LOCUS       V024677                10071 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024677
VERSION     V024677
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..4751
                     /label="MYBBP1A(NM_001105538)"
                     /note="MYBBP1A(NM_001105538)"
                     /gene="MYBBP1A"
     CDS             4758..4823
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4847..5435
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5501..5549
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5751..6179
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6193..6522
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6589..7383
                     /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    7559..7680
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7729..7745)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7753..7769
                     /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(7777..7807)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7822..7843
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8131..8719)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8890..9750)
                     /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(9751..9855)
                     /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 ccaccatgga gagccgggat cccgcccagc
      781 cgatgtcgcc tggagaagcg acgcagagtg gcgcccggcc tgccgaccgc tatggcctat
      841 tgaagcacag tcgcgagttc ttggacttct tctgggacat tgcgaagcct gagcaggaga
      901 cgcgacttgc ggccacggag aagctgctgg agtatctgcg tggcaggccg aaggggtccg
      961 agatgaaata tgccctgaag cgtctaatca cgggactcgg ggtcgggcga gaaacagccc
     1021 ggccctgcta cagtttggcc ctggcacagc tgttacagtc ttttgaagac ctccccttgt
     1081 gcagcatcct gcagcagata caagaaaaat atgacctgca tcaggtgaag aaggcaatgc
     1141 tgagacctgc tctctttgca aacctgtttg gagtgctcgc cctctttcag tcaggtcggc
     1201 tggtgaagga ccaggaggca ctgatgaagt cggtgaagct gctgcaggcc ctggcccagt
     1261 accaaaacca cttgcaggag cagccccgga aggccctggt ggacatcctc tccgaggtct
     1321 cgaaggccac attgcaggag atcctgccgg aggtcctcaa agccgacttg aatataatac
     1381 tcagctcccc tgaacagcta gagctcttcc tcctggccca gcagaaggtg ccctccaagc
     1441 tcaagaagct ggtgggatcc gtgaacctat tctcagatga gaatgtcccc aggctggtga
     1501 atgtgctgaa gatggccgcc tcctctgtga agaaggaccg caagctgccc gccattgctc
     1561 tggacctgct ccgcctggca ctcaaggaag acaagttccc acggttctgg aaggaggtgg
     1621 tggaacaagg gctgctgaag atgcagttct ggccagccag ctacctgtgt ttccgcctgc
     1681 tgggcgcggc cctgcccctg ctgaccaagg agcagctgca cctggtgatg cagggagacg
     1741 tgatccgcca ttacggggag cacgtgtgca ctgctaagct cccaaagcag ttcaagtttg
     1801 ccccagagat ggacgattac gtgggcacct tcctagaggg gtgccaggat gaccctgagc
     1861 ggcagctggc cgtgctagtg gccttctcat ctgtcaccaa ccaaggcctc cctgtcacgc
     1921 ctactttctg gcgggtcgtg cggttcctga gccctccggc cctgcagggc tatgtggcct
     1981 ggctgcgggc catgtttctc cagccagacc tggactcctt ggttgacttc agcaccaaca
     2041 accagaagaa agcccaggat tcatcgctcc acatgcctga gcgagctgtg ttccggctga
     2101 ggaaatggat catctttcga ttggtgagca ttgtggacag cctgcacctg gagatggagg
     2161 aggccttgac tgagcaggtg gccaggtttt gtttgttcca ctcgttcttt gtcacaaaga
     2221 agcccacatc ccagatccct gagacaaagc acccgttctc cttccctttg gaaaaccagg
     2281 cccgagaggc tgtcagcagt gccttcttca gtctgttgca gaccctcagc acgcagttca
     2341 agcaggcacc gggccagacc cagggtgggc agccctggac ctaccacctg gtgcagttcg
     2401 cagacctcct gttgaatcac agccacaacg tgaccaccgt gacacccttc actgcgcagc
     2461 agcgccaggc ctgggaccgg atgctgcaga ctctgaagga gctggaggcc cactccgcag
     2521 aggccagggc tgctgccttc cagcaccttc tgctcctcgt gggcatccac ctcctcaagt
     2581 cccctgcaga gagctgtgac ctgctgggtg acatccagac ctgcatcagg aaaagtctgg
     2641 gagagaagcc ccgccggagc cgcaccaaga ccatcgaccc ccaggaaccc ccgtgggtag
     2701 aggtgctggt ggagatcttg ctggccctgt tggcccagcc cagccacctc atgcgccagg
     2761 tggcccggag cgtgtttggc cacatctgct cccacctgac cccgcgtgcc ctgcagctaa
     2821 ttctggatgt gctgaacccc gagaccagtg aggatgagaa tgaccgtgtg gtggtgacgg
     2881 acgattctga tgagcggcgg ctgaagggtg cagaggacaa gagcgaggaa ggtgaggaca
     2941 acagaagctc agagagtgaa gaggagagcg agggggagga gagcgaggag gaggagcgcg
     3001 acggggacgt ggatcagggc ttccgggaac agctgatgac cgtgctgcag gctgggaagg
     3061 cgctgggtgg agaggacagt gagaacgagg aggagctggg ggatgaggcc atgatggccc
     3121 tggaccagag cctcgccagc ctctttgccg agcagaagct gcgtatccag gcccggcgag
     3181 acgagaagaa caagctgcag aaggagaagg ctctgcggcg cgacttccag atccgggtgc
     3241 tggacctggt ggaggtgcta gtgaccaagc agcccgagaa tgccctggtc ctggagctgc
     3301 tggagccgct gctgagcatc atccggcgca gcctgcgcag cagcagctcc aaacaggagc
     3361 aggaccttct gcacaagacg gcgcgcatct tcacgcacca cctgtgccgt gcccggcgct
     3421 actgccacga cttgggtgag cgcgcagggg ccctgcacgc ccaggtggag cggttggtgc
     3481 agcaggctgg ccgccagccc gactccccca ccgccctcta ccacttcaac gcctctctct
     3541 acctgctccg ggtcttgaag ggcaacactg ctgagggctg cgtgcatgag acacaggaga
     3601 agcagaaagc tggcactgac cccagccaca tgcccacggg cccgcaggct gccagctgct
     3661 tggacttgaa cctggtgacc cgggtgtact cgacagcact gagctccttc ctgaccaagc
     3721 gcaacagccc cctcacagtt cccatgttcc tcagcctctt ctcccggcac ccggtgctct
     3781 gtcagagcct gctccccatc ctggtccagc atatcacggg cccggtgcgg ccccgtcatc
     3841 aggcctgcct gctgctccag aagaccctgt ccatgcggga ggtgaggtcg tgctttgagg
     3901 accccgagtg gaagcagctg atgggccagg tcctagcaaa ggtcaccgag aacttgcgcg
     3961 tgctggggga ggcgcagacc aaggcgcagc atcagcaggc actgtcctcc ctggagctgc
     4021 tcaacgttct cttcaggacc tgcaaacatg agaagctgac cttggacctg acggtgctcc
     4081 tgggtgtgct gcaggggcaa cagcagagcc tacagcaggg ggcacactcc accggctcca
     4141 gccgcctgca cgacctctac tggcaggcca tgaaaaccct gggagtccag cgccccaagt
     4201 tggagaagaa ggatgccaag gagatcccca gtgccaccca gagccccatc agtaagaagc
     4261 ggaagaaaaa gggattcttg ccagagacga agaagcgcaa gaaacgcaag tcagaggatg
     4321 gcacgccagc ggaggatggc acacctgcag ccaccggcgg gagccagccc cccagcatgg
     4381 gcaggaagaa gaggaacagg acaaaggcta aggtcccagc ccaggcaaac gggacgccaa
     4441 ccaccaagag tccagcccct ggcgccccca cccggagccc cagcacccct gccaaatccc
     4501 caaaactgca gaagaaaaac cagaagccgt cccaggtgaa tggagctccc gggtccccca
     4561 cggaacctgc aggccaaaag cagcatcaga aggctcttcc caaaaagggg gtcttgggca
     4621 aatcaccact gtccgcgctg gcacggaaaa aggcaaggct gtctttggtc atcaggagtc
     4681 ccagcctgct tcagagtggg gccaagaaga aagcacagac tctgagattc acaatcagca
     4741 gctctaaaaa aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     4801 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     4861 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4921 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4981 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5041 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5101 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5161 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5221 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5281 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5341 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5401 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     5461 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5521 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5581 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5641 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5701 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     5761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5881 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     5941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6121 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     6181 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     6241 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     6301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     6361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6421 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     6481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     6541 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     6601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     7021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     7081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     7141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     7201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     7261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     7321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     7381 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     7441 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     7501 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     7561 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7621 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7681 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     7741 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7801 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7861 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     8041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     8101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     8161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     8221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     8281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     8341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     8401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     8461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     8521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     8581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     9001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     9061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     9121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     9181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     9241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     9301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     9361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9901 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     9961 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
    10021 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
