LOCUS       V027086                 8879 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027086
VERSION     V027086
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             950..4399
                     /label="CACNA2D2(NM_001174051)"
                     /note="CACNA2D2(NM_001174051)"
                     /gene="CACNA2D2"
     promoter        complement(4431..4449)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    4475..4699
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4745..5173
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5187..5517
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5584..6378
                     /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    6552..6673
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6722..6738)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6746..6762
                     /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(6770..6800)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6815..6836
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7124..7712)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7883..8743)
                     /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(8744..8848)
                     /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 ccactagtaa cggccgccag tgtgctggaa ttcgccacca tggctgtgcc
      961 tgctaggacc tgcggagctt ctagacccgg acctgctaga accgccagac cttggcctgg
     1021 atgcggacct catcccggac ctggaacaag aaggcccacc tctggccctc ctagacctct
     1081 gtggctgctg ctgcctctgc tgcctctgct ggctgctcct ggagcttctg cttactcttt
     1141 cccccagcag cacaccatgc agcactgggc tagacgtctg gagcaggagg tcgacggcgt
     1201 gatgcggatt tttggaggcg tccagcagct ccgtgagatt tacaaggaca accggaacct
     1261 gttcgaggta caggagaatg agcctcagaa gttggtggag aaggtggcag gggacattga
     1321 gagccttctg gacaggaagg tgcaggccct gaagagactg gctgatgctg cagagaactt
     1381 ccagaaagca caccgctggc aggacaacat caaggaggaa gacatcgtgt actatgacgc
     1441 caaggctgac gctgagctgg acgaccctga gagtgaggat gtggaaaggg ggtctaaggc
     1501 cagcacccta aggctggact tcatcgagga cccaaacttc aagaacaagg tcaactattc
     1561 atacgcggct gtacagatcc ctacggacat ctacaaaggc tccactgtca tcctcaatga
     1621 gctcaactgg acagaggccc tggagaatgt gttcatggaa aaccgcagac aagaccccac
     1681 actgctgtgg caggtcttcg gcagcgccac aggagtcact cgctactacc cggccacccc
     1741 gtggcgagcc cccaagaaga tcgacctgta cgatgtccga aggagaccct ggtatatcca
     1801 gggggcctcg tcacccaaag acatggtcat catcgtggat gtgagtggca gtgtgagcgg
     1861 cctgaccctg aagctgatga agacatctgt ctgcgagatg ctggacacgc tgtctgatga
     1921 tgactatgtg aatgtggcct cgttcaacga gaaggcacag cctgtgtcat gcttcacaca
     1981 cctggtgcag gccaatgtgc gcaacaagaa ggtgttcaag gaagctgtgc agggcatggt
     2041 ggccaagggc accacaggct acaaggccgg ctttgagtat gcctttgacc agctgcagaa
     2101 ctccaacatc actcgggcca actgcaacaa gatgatcatg atgttcacgg atggtggtga
     2161 ggaccgcgtg caggacgtct ttgagaagta caattggcca aaccggacgg tgcgcgtgtt
     2221 tactttctcc gtggggcagc ataactatga cgtcacaccg ctgcagtgga tggcctgtgc
     2281 caacaaaggc tactattttg agatcccttc catcggagcc atccgcatca acacacagga
     2341 atatctagat gtgttgggca ggcccatggt gctggcaggc aaggaggcca agcaggtgca
     2401 gtggaccaac gtgtatgagg atgcactggg actggggttg gtggtaacag ggaccctccc
     2461 tgttttcaac ctgacacagg atggccctgg ggaaaagaag aaccagctga tcctgggcgt
     2521 gatgggcatt gacgtggctc tgaatgacat caagaggctg acccccaact acacgcttgg
     2581 agccaacggc tatgtgtttg ccattgacct gaacggctac gtgttgctgc accccaatct
     2641 caagccccag accaccaact tccgggagcc tgtgactctg gacttcctgg atgcggagct
     2701 agaggatgag aacaaggaag agatccgtcg gagcatgatt gatggcaaca agggccacaa
     2761 gcagatcaga acgttggtca agtccctgga tgagaggtac atagatgagg tgacacggaa
     2821 ctacacctgg gtgcctataa ggagcactaa ctacagcctg gggctggtgc tcccacccta
     2881 cagcaccttc tacctccaag ccaatctcag tgaccagatc ctgcaggtca agttgccaat
     2941 cagcaaactg aaggattttg agttcctgct ccccagcagc tttgagtctg aaggacacgt
     3001 tttcattgct cccagagagt actgcaagga cctgaatgcc tcagacaaca acaccgagtt
     3061 cctgaaaaac tttattgagc tcatggagaa agtgactcca gactccaagc agtgcaacaa
     3121 cttccttctg cacaacctga tcttggacac gggcatcacg cagcagctgg tagagcgtgt
     3181 gtggagggac caggatctca acacgtacag cctactggcc gtgttcgctg ccacagacgg
     3241 tggcatcacc cgagtcttcc ccaacaaggc agctgaggac tggacagaga accctgagcc
     3301 cttcaatgcc agcttctacc gccgcagcct ggataaccac ggttatgtct tcaagccccc
     3361 acaccaggat gccctgttaa ggccgctgga gctggagaat gacactgtgg gcatcctcgt
     3421 cagcacagct gtggagctca gcctaggcag gcgcacactg aggccagcag tggtgggcgt
     3481 caagctggac ctagaggctt gggctgagaa gttcaaggtg ctagccagca accgtaccca
     3541 ccaagaccag cctcagaagt gcggccccaa cagccactgt gagatggact gcgaggttaa
     3601 caatgaggac ttactctgtg tcctcattga tgatggagga ttcctggtgc tgtcaaacca
     3661 gaaccatcag tgggaccagg tgggcaggtt cttcagtgag gtggatgcca acctgatgct
     3721 ggcactctac aataactcct tctacacccg caaggagtcc tatgactatc aggcagcctg
     3781 tgcccctcag ccccctggca acctgggtgc tgcaccccgg ggtgtctttg tgcccaccgt
     3841 tgcagatttc cttaacctgg cctggtggac ctctgctgcc gcctggtccc tgttccagca
     3901 gcttctctac ggcctcatct accacagctg gttccaagca gaccccgcgg aggccgaggg
     3961 gagccccgag acgcgcgaga gcagctgcgt catgaaacag acccagtact acttcggctc
     4021 ggtaaacgcc tcctacaacg ccatcatcga ctgcggaaac tgctccaggc tgttccacgc
     4081 gcagagactg accaacacca atcttctctt tgtggtggcc gagaagccgc tgtgcagcca
     4141 gtgcgaggct ggccggctgc tgcagaagga gacgcactcg gacggcccgg agcagtgtga
     4201 gctagtgcag agaccgcgat accggagagg cccgcacatc tgcttcgact acaacgcgac
     4261 agaagatacc tcagactgtg gccgcggggc ctccttcccg ccgtcgctgg gcgtcctggt
     4321 ctccctgcaa ctgctgctcc tcctgggcct gccgccccgg ccgcagcctc aagtcctcgt
     4381 ccacgcctct cgccgcctct gaactcgagc atgcatctag agggccctat tctatagtgt
     4441 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4501 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4561 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4621 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4681 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4741 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4801 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4861 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4921 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4981 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5041 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5101 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5161 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5221 ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5281 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5341 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5401 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5461 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5521 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5581 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5641 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5701 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5761 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5821 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5881 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5941 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6001 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6061 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6121 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6181 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6241 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6301 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6361 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6421 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6481 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6541 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6601 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6661 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6721 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6781 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6841 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6961 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7021 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7081 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     7321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7561 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7861 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7921 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7981 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8041 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8101 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8161 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8221 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8281 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8341 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8401 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8461 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8521 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8581 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8641 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8701 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8761 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8821 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
