LOCUS       V027724                 6852 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027724
VERSION     V027724
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             919..2340
                     /label="CBLC(NM_012116)"
                     /note="CBLC(NM_012116)"
                     /gene="CBLC"
     CDS             2347..2373
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        complement(2404..2422)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2448..2672
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2718..3146
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3160..3490
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3557..4351
                     /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    4525..4646
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4695..4711)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4719..4735
                     /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(4743..4773)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4788..4809
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5097..5685)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5856..6716)
                     /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(6717..6821)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccgccaccat ggctctggcg gtggccccgt gggggcgaca gtgggaagag
      961 gcccgcgccc tgggccgggc agtcaggatg ctgcagcgcc tagaagagca atgcgtcgac
     1021 ccccggctgt ccgtgagtcc cccttcgctg cgggacctgc tgccccgcac agcgcagctg
     1081 cttcgagagg tggcccattc tcggcgggcg gccggcggag gcggccccgg gggtcccggc
     1141 ggctctgggg actttctact catctacctg gccaatctgg aggccaagag caggcaggtg
     1201 gccgcgctgc tgcctccccg gggccgaagg agtgccaacg acgagctctt ccgggcgggc
     1261 tccagactca ggcgacagct ggccaagctg gccatcatct tcagccacat gcacgcagag
     1321 ctgcacgcac tcttccccgg gggaaagtac tgtggacaca tgtaccagct caccaaggcc
     1381 cccgcccaca ccttctggag ggaaagttgc ggagcccggt gtgtgctgcc ctgggctgag
     1441 tttgagtccc tcctgggcac ctgccaccct gtggaaccag gctgcacagc cctggccttg
     1501 cgcaccacca ttgacctcac ctgcagcggg cacgtgtcca tcttcgagtt cgacgtcttc
     1561 accaggctct ttcagccatg gccaacactc ctcaagaact ggcagctcct ggcagtcaac
     1621 cacccaggct acatggcctt cctcacctat gatgaggtcc aagagcgtct gcaggcctgc
     1681 agggacaagc caggcagtta catcttccgg cccagctgta ctcgcctggg gcagtgggcc
     1741 atcggctatg tgagctcaga tggcagcatc ctgcagacca tccctgccaa caaacccctg
     1801 tcccaggtgc tcctggaggg acagaaggac ggcttctacc tctacccaga tggaaagacc
     1861 cacaacccag acctgactga gctcggccag gcagaacccc agcagcgcat ccacgtgtca
     1921 gaggagcagc tgcagctcta ctgggccatg gactccacat ttgagctctg caagatctgt
     1981 gctgagagca acaaggatgt gaagattgag ccgtgcgggc acctgctctg cagctgctgc
     2041 ctggctgcct ggcagcactc ggacagccag acctgcccct tctgccgctg cgagatcaag
     2101 ggctgggagg ccgtgagtat ctaccagttc cacggtcagg ctactgctga ggactcaggg
     2161 aacagcagtg accaggaagg cagggagttg gagctggggc aggtgcccct ttcggctcct
     2221 ccattgcccc cacggccaga tctgcccccc aggaagccca gaaatgccca gccgaaagtg
     2281 agactcctaa aggggaactc ccctccagct gcgctgggac cccaggaccc tgccccggcc
     2341 gaattctacc catacgacgt accagattac gcttgactcg agcatgcatc tagagggccc
     2401 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     2461 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2521 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2581 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2641 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2701 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3061 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3121 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3181 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     3241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3421 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3481 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3541 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3601 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3661 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3721 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3781 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3841 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3901 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3961 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4021 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4081 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4141 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4201 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4261 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4321 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4381 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4441 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4501 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4561 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4621 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4681 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4741 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4801 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4861 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5281 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5521 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6841 ccacctgacg tc
//
