LOCUS       V037277                 6893 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037277
VERSION     V037277
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"
     CDS             908..2329
                     /label="CBLC(NM_012116)"
                     /note="CBLC(NM_012116)"
                     /gene="CBLC"
     CDS             2336..2425
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2460..2684
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2730..3158
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3172..3501
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3568..4362
                     /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    4536..4657
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4706..4722)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4730..4746
                     /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(4754..4784)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4799..4820
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5108..5693)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5864..6724)
                     /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(6725..6829)
                     /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 cttggtaccg agctcggatc
      901 cgccaccatg gctctggcgg tggccccgtg ggggcgacag tgggaagagg cccgcgccct
      961 gggccgggca gtcaggatgc tgcagcgcct agaagagcaa tgcgtcgacc cccggctgtc
     1021 cgtgagtccc ccttcgctgc gggacctgct gccccgcaca gcgcagctgc ttcgagaggt
     1081 ggcccattct cggcgggcgg ccggcggagg cggccccggg ggtcccggcg gctctgggga
     1141 ctttctactc atctacctgg ccaatctgga ggccaagagc aggcaggtgg ccgcgctgct
     1201 gcctccccgg ggccgaagga gtgccaacga cgagctcttc cgggcgggct ccagactcag
     1261 gcgacagctg gccaagctgg ccatcatctt cagccacatg cacgcagagc tgcacgcact
     1321 cttccccggg ggaaagtact gtggacacat gtaccagctc accaaggccc ccgcccacac
     1381 cttctggagg gaaagttgcg gagcccggtg tgtgctgccc tgggctgagt ttgagtccct
     1441 cctgggcacc tgccaccctg tggaaccagg ctgcacagcc ctggccttgc gcaccaccat
     1501 tgacctcacc tgcagcgggc acgtgtccat cttcgagttc gacgtcttca ccaggctctt
     1561 tcagccatgg ccaacactcc tcaagaactg gcagctcctg gcagtcaacc acccaggcta
     1621 catggccttc ctcacctatg atgaggtcca agagcgtctg caggcctgca gggacaagcc
     1681 aggcagttac atcttccggc ccagctgtac tcgcctgggg cagtgggcca tcggctatgt
     1741 gagctcagat ggcagcatcc tgcagaccat ccctgccaac aaacccctgt cccaggtgct
     1801 cctggaggga cagaaggacg gcttctacct ctacccagat ggaaagaccc acaacccaga
     1861 cctgactgag ctcggccagg cagaacccca gcagcgcatc cacgtgtcag aggagcagct
     1921 gcagctctac tgggccatgg actccacatt tgagctctgc aagatctgtg ctgagagcaa
     1981 caaggatgtg aagattgagc cgtgcgggca cctgctctgc agctgctgcc tggctgcctg
     2041 gcagcactcg gacagccaga cctgcccctt ctgccgctgc gagatcaagg gctgggaggc
     2101 cgtgagtatc taccagttcc acggtcaggc tactgctgag gactcaggga acagcagtga
     2161 ccaggaaggc agggagttgg agctggggca ggtgcccctt tcggctcctc cattgccccc
     2221 acggccagat ctgcccccca ggaagcccag aaatgcccag ccgaaagtga gactcctaaa
     2281 ggggaactcc cctccagctg cgctgggacc ccaggaccct gccccggccc tcgaggaaca
     2341 aaaactaata tcagaggaag atttggaaca gaaactaatc agtgaagaag atttagagca
     2401 aaagttaatt tccgaggagg acttatgagg gcccgtttaa acccgctgat cagcctcgac
     2461 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2521 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2581 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2641 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2701 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2761 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2821 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2881 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2941 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3001 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3061 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3121 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3181 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3241 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3361 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3421 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3481 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3541 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3601 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3661 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3721 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3781 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3841 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3901 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3961 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4021 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4081 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4141 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4201 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4261 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4321 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4381 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4441 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4501 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4681 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4741 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4801 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4861 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5521 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5641 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5821 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5881 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5941 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6001 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6061 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6121 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6181 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6241 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6301 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6361 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6421 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6481 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6541 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6601 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6661 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6841 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
