LOCUS       V036147                 9507 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036147
VERSION     V036147
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4187
                     /label="ADCY3(NM_001320613)"
                     /note="ADCY3(NM_001320613)"
                     /gene="ADCY3"
     CDS             4194..4259
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4283..4871
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4937..4985
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5187..5615
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5629..5958
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6025..6819
                     /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    6995..7116
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7165..7181)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7189..7205
                     /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(7213..7243)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7258..7279
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7567..8155)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8326..9186)
                     /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(9187..9291)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcc gaggaaccag ggcttctccg
      781 agcccgaata ctcggccgag tactcagccg agtactccgt cagcctgccc tccgaccctg
      841 accgcggggt gggccggacc catgaaatct cggtccggaa ctcgggctcc tgcctgtgcc
      901 tgcctcgctt catgcggctg actttcgtgc cggagtcctt ggagaacctc taccagacct
      961 acttcaaaag gcagcgccac gagaccctgc tggtgctggt ggtctttgca gccctctttg
     1021 actgctacgt ggtggtcatg tgtgctgtgg tcttctccag cgacaagctg gcttccctcg
     1081 ccgtggctgg aattggactg gtgttggaca tcatcctctt cgtgctctgc aaaaaggggc
     1141 tgctcccgga ccgggtcacc cgcagagtgc tgccctacgt gctgtggctg ctcataaccg
     1201 cccagatctt ctcctacctg ggcctgaact tcgcgcgtgc ccacgcggct agtgacacgg
     1261 tgggctggca ggtcttcttt gtcttctcct tcttcatcac gctgcccctc agcctcagcc
     1321 ccatcgtgat catctccgtg gtctcctgtg tggtgcacac gttggtcctg ggggtcaccg
     1381 tggcccagca gcagcaggag gagctcaagg ggatgcagct gctgcgggag atcctggcca
     1441 acgtcttcct ctacctgtgc gccatcgctg tgggcatcat gtcctactac atggctgacc
     1501 gcaagcaccg caaggccttc ctggaggccc gccagtcgct ggaggtgaag atgaacctgg
     1561 aagagcagag ccagcagcag gagaacctca tgctttccat cctgcccaag cacgtggctg
     1621 acgagatgct gaaagacatg aagaaagacg agagccagaa ggaccagcag cagttcaaca
     1681 ccatgtacat gtaccgtcac gagaacgtca gcatcctctt tgccgacatc gtgggcttta
     1741 cccagctgtc ttctgcctgc agtgcccagg agcttgtgaa gctgctcaac gagctctttg
     1801 cccgctttga caagctggca gctaaatacc accagctgcg gattaagatc ctgggcgact
     1861 gctactactg catctgcggc ttgcccgact accgggagga ccacgccgtc tgctccatcc
     1921 tcatggggct ggccatggtg gaggccatct cgtatgtgcg ggagaagacc aagactgggg
     1981 tggacatgcg tgtgggggtg cacacgggca ccgtgctggg gggcgtcctg ggccagaagc
     2041 gctggcagta cgacgtgtgg tcgactgatg tcactgtagc caacaagatg gaggccggcg
     2101 gcatccctgg gcgcgtgcac atctcccaga gcaccatgga ctgcctgaaa ggggagtttg
     2161 atgtggagcc aggcgatggg ggcagccgct gtgattacct agaagagaag ggtattgaaa
     2221 cctacctcat cattgcctcc aagccagagg tgaagaaaac agccacccag aatggcctca
     2281 atggctcggc cctgcccaat ggagcaccag cttcctcaaa gtccagctcc cctgccctca
     2341 ttgagaccaa ggagcccaac gggagtgccc acagcagtgg gtccacgtcg gagaagcccg
     2401 aggagcagga tgcccaggcc gacaacccct cattccccaa cccacgccgg aggctgcgcc
     2461 tgcaggacct ggctgaccga gtggtggatg cctctgaaga tgagcacgag ctcaaccagc
     2521 tgctcaacga ggccctgctt gagcgagagt ccgcccaagt agtaaagaag agaaacacct
     2581 tcctcttgtc catgcggttc atggaccccg agatggaaac ccgctactcg gtggagaagg
     2641 agaagcagag tggggctgcc ttcagctgct cctgcgtcgt cctgctctgc acggccctgg
     2701 tcgagatact catcgacccc tggctaatga caaactatgt gaccttcatg gtgggggaga
     2761 ttctgctcct catcctgacc atctgctccc tggctgccat ctttccccgg gcctttccta
     2821 agaagcttgt ggccttctca acttggattg accggacccg ctgggccagg aacacctggg
     2881 ccatgctcgc catcttcatc ctggtgatgg caaatgtcgt ggacatgctc agctgtctcc
     2941 agtactacac gggacccagc aatgcaacgg cagggatgga aacggagggc agctgcctgg
     3001 agaaccccaa gtattacaac tatgtggccg tgctgtccct catcgccacc atcatgctgg
     3061 tgcaggtcag ccacatggtg aagctcacgc tcatgctgct cgtcgcaggc gccgtggcca
     3121 ccatcaacct ctatgcctgg cgtcccgtct ttgatgaata cgaccacaag cgttttcggg
     3181 agcacgactt acctatggtg gccttagagc agatgcaagg attcaaccct gggctcaatg
     3241 gcactgacag gctgcccctg gtgccttcca agtactctat gacggtgatg gtgttcctca
     3301 tgatgctcag cttctactac ttctcccgcc acgtagaaaa actggcacgg acacttttct
     3361 tgtggaagat tgaggtccac gaccagaagg aacgtgtcta tgagatgcga cgctggaacg
     3421 aggccttggt caccaacatg ttgcctgagc acgtggcacg ccatttcctg gggtccaaga
     3481 agagagatga ggagctgtat agccagacgt atgatgagat tggagtcatg tttgcctccc
     3541 tgcccaactt tgctgacttc tacacagagg agagcatcaa caatggtggt attgagtgtc
     3601 tgcgtttcct caatgaaatc atctcagatt ttgactctct cctggacaat cccaagttcc
     3661 gggtgatcac caagatcaaa accattggca gcacgtatat ggcggcttca ggagtcaccc
     3721 ccgatgtcaa caccaatggc tttgccagct ccaacaagga agacaagtcc gagagagagc
     3781 gctggcagca cctggctgac ctggccgact tcgcgctggc catgaaggat acgctcacca
     3841 acatcaacaa ccagtccttc aataacttca tgctgcgcat aggcatgaac aaaggcgggg
     3901 ttctggctgg ggtcatcgga gcccggaaac cacactacga catctggggc aatacagtca
     3961 atgtagccag caggatggag tccacggggg tcatgggcaa cattcaggtg gtagaagaaa
     4021 cccaagtcat cctccgagag tacggcttcc gctttgtgag gcgaggcccc atctttgtga
     4081 aggggaaggg ggagctgctg accttcttct tgaaggggcg ggataagcta gccaccttcc
     4141 ccaatggccc ctctgtcaca ctgccccacc aggtggtgga caactccgga tccgactaca
     4201 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     4261 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4321 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4381 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4441 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4501 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4561 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4621 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     4681 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4741 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4801 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4861 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     4921 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     4981 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5041 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5101 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     5161 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     5221 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5281 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5341 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5401 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5461 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5521 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     5581 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     5641 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5701 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5761 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5821 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5881 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5941 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     6001 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6061 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6121 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6181 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6241 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6301 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6361 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6421 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6481 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6541 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6601 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6661 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6721 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6781 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     6841 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6901 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6961 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     7021 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7081 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     7141 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7201 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7261 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7321 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7381 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7441 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7741 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7981 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     8041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8281 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8341 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8401 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8461 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8521 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8581 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8641 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8701 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8761 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8821 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8881 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8941 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9001 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9061 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9121 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9181 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9301 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     9361 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     9421 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     9481 gacaattgca tgaagaatct gcttagg
//
