LOCUS       V018511                 7181 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018511
VERSION     V018511
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..2701
                     /label="CACNB1(NM_000723)"
                     /note="CACNB1(NM_000723)"
                     /gene="CACNB1"
     polyA_signal    2748..2972
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3018..3446
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3460..3789
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3856..4650
                     /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    4824..4945
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4994..5010)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5018..5034
                     /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(5042..5072)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5087..5108
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5396..5981)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6152..7012)
                     /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(7013..7117)
                     /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 gtccagaaga ccagcatgtc ccggggccct tacccaccct cccaggagat
      961 ccccatggag gtcttcgacc ccagcccgca gggcaaatac agcaagagga aagggcgatt
     1021 caaacggtca gatgggagca cgtcctcgga taccacatcc aacagctttg tccgccaggg
     1081 ctcagcggag tcctacacca gccgtccatc agactctgat gtatctctgg aggaggaccg
     1141 ggaagcctta aggaaggaag cagagcgcca ggcattagcg cagctcgaga aggccaagac
     1201 caagccagtg gcatttgctg tgcggacaaa tgttggctac aatccgtctc caggggatga
     1261 ggtgcctgtg cagggagtgg ccatcacctt cgagcccaaa gacttcctgc acatcaagga
     1321 gaaatacaat aatgactggt ggatcgggcg gctggtgaag gagggctgtg aggttggctt
     1381 cattcccagc cccgtcaaac tggacagcct tcgcctgctg caggaacaga agctgcgcca
     1441 gaaccgcctc ggctccagca aatcaggcga taactccagt tccagtctgg gagatgtggt
     1501 gactggcacc cgccgcccca caccccctgc cagtgccaaa cagaagcaga agtcgacaga
     1561 gcatgtgccc ccctatgacg tggtgccttc catgaggccc atcatcctgg tgggaccgtc
     1621 gctcaagggc tacgaggtta cagacatgat gcagaaagct ttatttgact tcttgaagca
     1681 tcggtttgat ggcaggatct ccatcactcg tgtgacggca gatatttccc tggctaagcg
     1741 ctcagttctc aacaacccca gcaaacacat catcattgag cgctccaaca cacgctccag
     1801 cctggctgag gtgcagagtg aaatcgagcg aatcttcgag ctggcccgga cccttcagtt
     1861 ggtcgctctg gatgctgaca ccatcaatca cccagcccag ctgtccaaga cctcgctggc
     1921 ccccatcatt gtttacatca agatcacctc tcccaaggta cttcaaaggc tcatcaagtc
     1981 ccgaggaaag tctcagtcca aacacctcaa tgtccaaata gcggcctcgg aaaagctggc
     2041 acagtgcccc cctgaaatgt ttgacatcat cctggatgag aaccaattgg aggatgcctg
     2101 cgagcatctg gcggagtact tggaagccta ttggaaggcc acacacccgc ccagcagcac
     2161 gccacccaat ccgctgctga accgcaccat ggctaccgca gccctggctg ccagccctgc
     2221 ccctgtctcc aacctccagg gaccctacct tgcttccggg gaccagccac tggaacgggc
     2281 caccggggag cacgccagca tgcacgagta cccaggggag ctgggccagc ccccaggcct
     2341 ttaccccagc agccacccac caggccgggc aggcacgcta cgggcactgt cccgccaaga
     2401 cacttttgat gccgacaccc ccggcagccg aaactctgcc tacacggagc tgggagactc
     2461 atgtgtggac atggagactg acccctcaga ggggccaggg cttggagacc ctgcaggggg
     2521 cggcacgccc ccagcccgac agggatcctg ggaggacgag gaagaagact atgaggaaga
     2581 gctgaccgac aaccggaacc ggggccggaa taaggcccgc tactgcgctg agggtggggg
     2641 tccagttttg gggcgcaaca agaatgagct ggagggctgg ggacgaggcg tctacattcg
     2701 ctgactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2761 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2821 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2881 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2941 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3001 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3061 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3121 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3181 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3241 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3301 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3361 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3421 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3661 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3721 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3781 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3841 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3901 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3961 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4021 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4081 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4141 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4201 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4261 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4321 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4381 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4441 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4501 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4561 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4621 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4681 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4741 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4801 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4861 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4921 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4981 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5041 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5101 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5161 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     5941 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6001 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6061 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6121 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6181 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6241 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6301 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6361 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6421 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6481 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6541 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6601 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6661 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6721 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6781 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6841 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6901 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6961 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7021 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7141 ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
