LOCUS       V042928                 8381 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042928
VERSION     V042928
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..3871
                     /label="Bicc1(NM_031397)"
                     /note="Bicc1(NM_031397)"
                     /gene="Bicc1"
     CDS             3878..3943
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3981..4205
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4251..4679
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4693..5022
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5089..5883
                     /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    6057..6178
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6227..6243)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6251..6267
                     /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(6275..6305)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6320..6341
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6629..7214)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7385..8245)
                     /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(8246..8350)
                     /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 atggcctcgc agagcgagcc
      961 gggctacctg gcggcggcgc agtcggaccc cggctccaac agcgagcgca gcaccgactc
     1021 gccggtggcc ggctccgagg acgatctggt ggccgcggcg cccctcttgc acagcccgga
     1081 gtggagcgag gagcgcttcc gcgtggacag gaagaaactc gaggccatgc tccaagctgc
     1141 agctgaagga aaaggccgaa gtggggaaga cttttttcag aagatcatgg aggagacaaa
     1201 cacgcagatt gcatggccgt ccaaactgaa gatcggggct aaatccaaga aagatcccca
     1261 catcaaggtt tctgggaaga aagaggatgt gaaggaagcc aaagaaatga tcatgtctgt
     1321 cttagacaca aaaagcaacc gcgtcacatt gaagatggat gtctcgcaca cggagcactc
     1381 ccacgtcatc ggcaagggtg gtaacaacat taaaaaggtc atggaagaca cgggctgcca
     1441 catccacttc ccagactcca acaggaacaa ccaggcagag aagagtaacc aggtgtctat
     1501 agcaggacag ccagcaggag tagaatcggc ccgagcaagg attcgggagc tgcttccttt
     1561 ggtgctgatg tttgagttac cgattgccgg gattctccag ccagtccccg atcccaacac
     1621 cccgtccatt cagcacatct cacaaaccta cagcgtttct gtgtccttta agcagaggtc
     1681 tcgaatgtat ggtgctacag tcacagtacg aggctctcag aataacacta atgctgtgaa
     1741 ggaaggaaca gccatgctgt tggaacacct tgcgggaagc ttggcctccg ccatccccgt
     1801 gagcacacaa ctggacatag cagcccagca tcacctcttc atgatgggcc ggaacgggag
     1861 caacgtcaaa cacatcatgc agaggacagg ggcgcagatt cactttcccg accccagcaa
     1921 tccacagaag aaatccaccg tctacctcca gggcaccatt gagtctgtct gcctagcaag
     1981 gcagtatctc atggggtgtc ttcctctggt gttgatgttt gatatgaagg aagacattga
     2041 agtggaccca caggtcatcg cacagctgat ggaacagctg gacgtcttta tcagtattaa
     2101 accaaagccc aaacagccga gcaagtctgt gattgtgaaa agtgttgagc gaaatgcctt
     2161 aaatatgtat gaagcaagga agtgtctcct cggacttgaa agcagtgggg tttccatagc
     2221 aaccagtcta tccccagcat cgtgccctgc cggcctggcc tgtcccagcc tggatatctt
     2281 agcttcggca ggcctcggac tcactggact aggtttattg gggcccacca cattgtcgct
     2341 caatacgtca gccaccccaa actcactcct gaatgctctc aacacttcgg tcagtccttt
     2401 gcaaagttca agttctggta ctcccagtcc tacactgtgg gcacccccaa tcgctaacac
     2461 tgcaagcgcc acaggtttct ctacgatacc acaccttatg cttccctcta ctgcccaggc
     2521 cacattaacc aatattttgc tgtctggagt gcccacatac gggcacacgg ctccatctcc
     2581 cccacctggc ttgactcctg ttgatgttca catcaacagc atgcagacag aaggcaaaaa
     2641 catctctgcg tctataaatg gacatgtgca gcctgcaaac atgaaatacg gtccgctgtc
     2701 cacttcatcg cttggggaaa aagtgctgag ttcgaatcat ggtgacccat ccatgcagac
     2761 agctgggccc gaacaggctt ctcctaaatc aaactcggtg gaaggctgca atgatgcctt
     2821 tgttgaagtg ggcatgcctc gaagtccctc ccattctgga aacgctggcg acttgaagca
     2881 gatgctgggt gcctccaagg tctcctgtgc caagcggcag acggttgagc tactgcaggg
     2941 cacgaagaac tcgcacctcc acggcactga cagactactc tcagaccctg aactgagcgc
     3001 cacagaaagt ccgcttgctg acaagaaggc cccggggagc gaacgtgcag ctgagagggc
     3061 agcagctgcc cagcagaaat cggagagggc ccgcctggcc tcgcagccaa catatgtcca
     3121 catgcaggca tttgattatg agcagaagaa actattagcc accaaagcga tgttaaagaa
     3181 gccagtggtg actgaggtca gaacacctac gaatacgtgg agtggcctgg gattctcaaa
     3241 gtccatgccg gcagaaacca ttaaggaact gaggagagcc aaccacgtat cctataagcc
     3301 cacgatgacc accgcctatg agggctcctc attgtccctc tcaaggtcca gcagtcgtga
     3361 gcacctggca agtggaagcg agtcagacaa ctggagagac cggaatggaa taggccccat
     3421 gggtcacagt gaattctcag caccaatcgg cagccccaag cgcaagcaga acaaatcaag
     3481 agagcactat ctaagcagca gcaactacat ggactgcatt tcctcgctga cgggaagcaa
     3541 tggctgtaac ctgaacagct gcttcaaagg ctccgacctc cccgagcttt tcagcaagct
     3601 gggcctaggc aaatacacgg atgtcttcca gcagcaagag atcgatcttc agacattcct
     3661 caccctcaca gatcaggatc tgaaggagct gggaatcaca acctttggtg cccgaaggaa
     3721 aatgctgctg gcaatctcag agctaagtaa aaaccgaaga aaactttttg aaccaccaaa
     3781 cgcatcatgc acctccttcc tggaaggcgg agccagtggg aggctgcctc gccagtatca
     3841 ttcagacatt gcgagcgtca gtggccgctg gctcgaggac tacaaagacc atgacggtga
     3901 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     3961 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4021 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4081 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4141 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4201 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4261 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4321 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4381 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4441 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4501 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4561 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4621 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4681 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4861 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4921 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4981 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5041 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5101 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5161 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5221 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5281 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5341 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5401 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5461 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5521 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5581 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5641 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5701 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5761 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5821 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5881 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5941 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6001 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6061 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6121 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6181 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6241 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6301 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6361 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6421 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7141 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8341 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
