LOCUS       V042945                 8372 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042945
VERSION     V042945
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..3862
                     /label="BICC1(NM_001080512)"
                     /note="BICC1(NM_001080512)"
                     /gene="BICC1"
     CDS             3869..3934
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3972..4196
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4242..4670
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4684..5013
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5080..5874
                     /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    6048..6169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6218..6234)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6242..6258
                     /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(6266..6296)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6311..6332
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6620..7205)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7376..8236)
                     /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(8237..8341)
                     /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 atggccgccc agggagagcc
      961 cggctacctg gcggcgcagt cggaccccgg ctccaacagc gagcgcagca ccgactcccc
     1021 agtgcccggc tccgaggacg acttggtcgc cggggcgacc ctgcacagcc cggagtggag
     1081 cgaggagcgc ttccgcgtgg acaggaagaa acttgaggcc atgttacaag ctgctgctga
     1141 agggaaaggc agaagtgggg aagacttttt tcaaaagatc atggaggaaa caaatacgca
     1201 gattgcttgg ccatcaaaac tgaagatcgg agccaaatcc aagaaagatc cccatattaa
     1261 ggtttctgga aagaaagaag atgttaaaga agccaaggaa atgatcatgt ctgtcttaga
     1321 cacaaaaagc aatcgagtca cactgaagat ggatgtttca catacagaac attcacatgt
     1381 aatcggcaaa ggtggcaaca atattaaaaa agtgatggaa gaaaccggat gccatatcca
     1441 ctttccagat tccaacagga ataaccaagc agaaaaaagc aaccaggtat ctatagcggg
     1501 acaaccagca ggagtagaat ctgcccgagt tagaattcgg gagctgcttc ctttggtgct
     1561 gatgtttgag ctaccaattg ctggaattct tcaaccggtt cctgatccta attccccctc
     1621 tattcagcat atatcacaaa cgtacaatat ttcagtatca tttaaacagc gttcccgaat
     1681 gtatggtgct actgtcatag tacgagggtc tcagaataac actagtgctg tgaaggaagg
     1741 aactgccatg ctgttagaac atcttgctgg gagcttagca tcagctattc ctgtgagcac
     1801 acaactagat attgcagctc aacatcatct ctttatgatg ggtcgaaatg ggagcaacat
     1861 caaacatatc atgcagagaa caggtgctca gatccacttt cctgatccca gtaatccaca
     1921 aaagaaatct accgtctacc tccagggcac cattgagtct gtctgtcttg caaggcaata
     1981 tctcatgggt tgtcttcctc ttgtgttgat gtttgatatg aaggaagaaa ttgaagtaga
     2041 tccacaattc attgcgcagt tgatggaaca gcttgatgtc ttcatcagta ttaaaccaaa
     2101 gcccaaacag ccaagcaagt ctgtgattgt gaaaagtgtt gagcgaaatg ccttaaatat
     2161 gtatgaagca aggaaatgtc tcctcggact tgaaagcagt ggggttacca tagcaaccag
     2221 tccatcccca gcatcctgcc ctgccggcct ggcatgtccc agcctggata tcttagcttc
     2281 agcaggcctt ggactcactg gactaggtct tttgggaccc accaccttat ctctgaacac
     2341 ttcaacaacc ccaaactcac tcttgaatgc tcttaatagc tcagtcagtc ctttgcaaag
     2401 tccaagttct ggtacaccca gccccacatt atgggcaccc ccacttgcta atacttcaag
     2461 tgccacaggt ttttctgcta taccacacct tatgattcca tctactgccc aagccacatt
     2521 aactaatatt ttgttgtctg gagtgcccac ctatgggcac acagctccat ctccccctcc
     2581 tggcttgact cctgttgatg tccatatcaa cagtatgcag accgaaggca aaaaaatctc
     2641 tgctgcttta aatggacatg cacagtctcc agatataaaa tatggtgcaa tatccacttc
     2701 atcacttgga gaaaaagtgc tgagtgcaaa tcacggggat ccgtccatcc agacaagtgg
     2761 gtctgagcag acatctccca aatcaagccc cactgaaggt tgtaatgatg cttttgttga
     2821 agtaggcatg cctcgaagtc cttcccattc tgggaatgct ggtgacttga aacagatgat
     2881 gtgtccctcc aaggtttcct gtgccaaaag gcagacagtg gaactattgc aaggcacgaa
     2941 aaactcacac ttacacagca ctgacaggtt gctctcagac cctgaactga gtgctaccga
     3001 aagccctttg gctgacaaga aggctccagg gagtgagcgc gctgcagaga gggcagcagc
     3061 tgcccagcaa aactccgaaa gggcccacct tgctccacgg tcatcatatg tcaacatgca
     3121 ggcatttgac tatgaacaga agaagctatt agccaccaaa gctatgttaa agaaaccagt
     3181 ggtgacggag gtcagaacgc ccacaaatac ctggagtggc ctgggttttt ctaaatccat
     3241 gccagctgaa actatcaagg agttgagaag ggccaatcat gtgtcctata agcccacaat
     3301 gacaaccact tatgagggct catccatgtc cctttcacgg tccaacagtc gtgagcactt
     3361 gggaggtgga agcgaatctg ataactggag agaccgaaat ggaattggac ctggaagtca
     3421 tagtgaattt gcagcttcta ttggcagccc taagcgtaaa caaaacaaat caacggaaca
     3481 ctatctcagc agtagcaatt acatggactg catttcctcg ctgacaggaa gcaatggctg
     3541 taacttaaat agctctttca aaggttctga cctccctgag ctcttcagca aactgggcct
     3601 gggcaaatac acagatgttt tccagcaaca agagatcgat cttcagacat tcctcactct
     3661 cacagatcag gatctgaagg agctgggaat aactactttt ggtgccagga ggaaaatgct
     3721 gcttgcaatt tcagaactaa ataaaaaccg aagaaagctt tttgaatcgc caaatgcacg
     3781 cacctctttc ctggaaggtg gagcgagtgg aaggctaccc cgtcagtatc actcagacat
     3841 tgctagtgtc agtggccgct ggctcgagga ctacaaagac catgacggtg attataaaga
     3901 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     3961 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4021 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4081 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4141 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4201 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4261 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4321 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4381 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4441 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4501 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4561 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4621 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4681 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4741 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4801 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4861 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4921 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4981 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5041 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5101 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5161 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5221 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5281 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5341 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5401 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5461 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5521 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5581 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5641 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5701 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5761 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5821 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5881 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5941 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6001 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     6061 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6121 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6181 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6241 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6301 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6361 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6421 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6481 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6541 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6601 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6661 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6721 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6781 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6841 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6901 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6961 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7021 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7081 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7141 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7201 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7261 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7321 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7381 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7441 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7501 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7561 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7621 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7681 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7741 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7801 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7861 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7921 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7981 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8041 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8101 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8161 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8221 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8281 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8341 gcacatttcc ccgaaaagtg ccacctgacg tc
//
