LOCUS       V022892                 6605 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022892
VERSION     V022892
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..2095
                     /label="BSG(NM_001728)"
                     /note="BSG(NM_001728)"
                     /gene="BSG"
     CDS             2102..2167
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2205..2429
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2475..2903
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2917..3246
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3313..4107
                     /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    4281..4402
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4451..4467)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4475..4491
                     /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(4499..4529)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4544..4565
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4853..5438)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5609..6469)
                     /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(6470..6574)
                     /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 atggcggctg cgctgttcgt
      961 gctgctggga ttcgcgctgc tgggcaccca cggagcctcc ggggctgccg gcttcgtcca
     1021 ggcgccgctg tcccagcaga ggtgggtggg gggcagtgtg gagctgcact gcgaggccgt
     1081 gggcagcccg gtgcccgaga tccagtggtg gtttgaaggg cagggtccca acgacacctg
     1141 ctcccagctc tgggacggcg cccggctgga ccgcgtccac atccacgcca cctaccacca
     1201 gcacgcggcc agcaccatct ccatcgacac gctcgtggag gaggacacgg gcacttacga
     1261 gtgccgggcc agcaacgacc cggatcgcaa ccacctgacc cgggcgccca gggtcaagtg
     1321 ggtccgcgcc caggcagtcg tgctagtcct ggaacccggc acagtcttca ctaccgtaga
     1381 agaccttggc tccaagatac tcctcacctg ctccttgaat gacagcgcca cagaggtcac
     1441 agggcaccgc tggctgaagg ggggcgtggt gctgaaggag gacgcgctgc ccggccagaa
     1501 aacggagttc aaggtggact ccgacgacca gtggggagag tactcctgcg tcttcctccc
     1561 cgagcccatg ggcacggcca acatccagct ccacgggcct cccagagtga aggctgtgaa
     1621 gtcgtcagaa cacatcaacg agggggagac ggccatgctg gtctgcaagt cagagtccgt
     1681 gccacctgtc actgactggg cctggtacaa gatcactgac tctgaggaca aggccctcat
     1741 gaacggctcc gagagcaggt tcttcgtgag ttcctcgcag ggccggtcag agctacacat
     1801 tgagaacctg aacatggagg ccgaccccgg ccagtaccgg tgcaacggca ccagctccaa
     1861 gggctccgac caggccatca tcacgctccg cgtgcgcagc cacctggccg ccctctggcc
     1921 cttcctgggc atcgtggctg aggtgctggt gctggtcacc atcatcttca tctacgagaa
     1981 gcgccggaag cccgaggacg tcctggatga tgacgacgcc ggctctgcac ccctgaagag
     2041 cagcgggcag caccagaatg acaaaggcaa gaacgtccgc cagaggaact cttccctcga
     2101 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2161 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     2221 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2281 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2341 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2401 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2461 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2521 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2581 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2641 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2701 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2761 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2821 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2881 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2941 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3001 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3061 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3121 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3181 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3241 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3301 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3361 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3421 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3481 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3541 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3601 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3661 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3721 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3781 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3841 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3901 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3961 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4021 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4081 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4141 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4201 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4261 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4321 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4381 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4441 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4501 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4561 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4621 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4681 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4741 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4801 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5041 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5281 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     5401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6601 acgtc
//
