LOCUS       V046479                 7137 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046479
VERSION     V046479
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..1817
                     /label="Gnao1(NM_010308)"
                     /note="Gnao1(NM_010308)"
                     /gene="Gnao1"
     CDS             1824..1889
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1913..2501
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2567..2615
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2817..3245
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3259..3588
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3655..4449
                     /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    4625..4746
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4795..4811)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4819..4835
                     /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(4843..4873)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4888..4909
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5197..5785)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5956..6816)
                     /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(6817..6921)
                     /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 ccaccatggg atgtacgctg agcgcagagg
      781 agagagccgc cctcgagcgg agcaaggcga ttgagaaaaa cctcaaagaa gatggcatca
      841 gcgccgccaa agacgtgaaa ttactcctgc tgggggctgg agaatcagga aaaagcacca
      901 ttgtgaagca gatgaagatc atccatgaag atggcttctc tggggaagac gtgaagcagt
      961 acaagcctgt ggtctacagc aacaccatcc agtctctggc ggccattgtc cgggccatgg
     1021 acactttggg cgtggagtat ggtgacaagg agaggaagac ggactccaag atggtgtgtg
     1081 acgtggtgag tcgtatggaa gacactgaac cgttctctgc agaacttctt tctgccatga
     1141 tgcgactctg gggcgactcg gggatccagg agtgcttcaa ccgatctcgg gagtatcagc
     1201 tcaatgactc tgccaaatac tacctggaca gcctggatcg gattggagcc ggtgactacc
     1261 agcccactga gcaggacatc ctccgaacca gagtcaaaac aactggcatc gtagaaaccc
     1321 acttcacctt caagaacctc cacttcaggc tgtttgacgt cgggggccag cgatctgaac
     1381 gcaagaagtg gatccactgc tttgaggatg tcacggccat catcttctgt gtcgcactca
     1441 gcggctatga ccaggtgctc cacgaggacg aaaccacgaa ccgcatgcac gagtctctca
     1501 tgctcttcga ctccatctgt aacaacaagt ttttcattga tacctccatc atcctcttcc
     1561 tcaacaagaa agacctcttt ggcgagaaga ttaagaagtc acccttgacc atctgctttc
     1621 ccgaataccc aggctccaac acctatgaag atgcagctgc ctacatccaa acacagtttg
     1681 aaagcaaaaa ccgctcaccc aacaaagaaa tttactgtca catgacttgt gccacagaca
     1741 cgaataatat ccaggtggta ttcgacgccg tcaccgacat catcattgcc aacaatctcc
     1801 ggggctgcgg cttgtacgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1861 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1921 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1981 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2041 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2101 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2161 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2221 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2281 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2341 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2401 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2461 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2521 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2581 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2641 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2701 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2761 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2821 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2881 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2941 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3001 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3061 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3121 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3181 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3241 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3301 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3421 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3481 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3541 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3601 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3661 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3721 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3781 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3841 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3901 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3961 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4021 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4081 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4141 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4201 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4261 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4321 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4381 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4441 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4501 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4561 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4621 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4741 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4801 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4861 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4921 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4981 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5041 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5101 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5161 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5881 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5941 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6001 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6061 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6121 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6181 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6241 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6301 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6361 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6421 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6481 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6541 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6601 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6661 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6721 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6781 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6841 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6901 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6961 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     7021 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7081 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
