LOCUS       V046281                 7878 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046281
VERSION     V046281
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..2558
                     /label="Ppp2r1b(NM_028614)"
                     /note="Ppp2r1b(NM_028614)"
                     /gene="Ppp2r1b"
     CDS             2565..2630
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2654..3242
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3308..3356
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3558..3986
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4000..4329
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4396..5190
                     /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    5366..5487
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5536..5552)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5560..5576
                     /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(5584..5614)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5629..5650
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5938..6526)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6697..7557)
                     /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(7558..7662)
                     /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 ccaccatggc gggcgcagca gggccagggt
      781 ccggtccggg ggcagcgggt ggagatggag acgattcgct ctacccgatc gcggttttaa
      841 tcgacgagct ccgcaatgag gacgtgcagc tccgtctcaa cagtattaag aagttatcaa
      901 ctatagctct agcactcggg gtagaaagga ctcgaacgga actgctgcct ttccttacag
      961 atactattta tgatgaagat gaggtgctgt tagctctcgc cgagcagctg ggaaatttca
     1021 ctggtttggt gggcggtcct gactttgccc actgtctgct gcctcctttg gaaagtttgg
     1081 ctacggtgga ggagactgtc gttcgagaca aggctgtgga gtcgctgagg cagatctcgc
     1141 aggagcacac tcctgttgcc cttgaagctc attttgtgcc tctggtgaag cgcctggcaa
     1201 gcggggactg gttcacgtcg cgcacgtctg catgtggctt gttcagcgtt tgctatccca
     1261 gagcttcaaa tgctgtcaag gcagagattc gacagcactt ccgttctctc tgctcggatg
     1321 atacaccaat ggtgcgacgt gctgctgctt ccaaattggg tgaatttgca aaagttttgg
     1381 aattagacag tgtgaaaact gaaattgtcc cattgttcac gaatttggct tcagatgaac
     1441 aggactctgt gcggctgcta gctgtggaag cctgcgtcag cattgcccag ttactgtctc
     1501 aggaggacct ggaagccttg gtaatgccta cacttcgaca agcagcagaa gataagtcct
     1561 ggcgggttcg atatatggtg gctgacaagt tttcagagct ccagaaagcg gtgggcccta
     1621 agattgcact cagcgacctc atccccgcct tccagagcct cctccgagac tgcgaagctg
     1681 aagtccgagc agccgctgcc cacaaagtaa gagagctttg tgagaacctg cccgctgaag
     1741 gtagagagac tgtaattatg aatcaaatcc tgccctatat aaaggaatta gtgtctgata
     1801 ccaaccaaca tgtcaagtca gcgctagctt ctgtaattat gggattatct acagttttgg
     1861 gcaaagagaa taccattgaa catcttctgc ctctattttt agctcagtta aaggacgagt
     1921 gtccagaagt tcgtttaaat atcatctcca atttggattg tgtgaatgaa gtgattggaa
     1981 tccgtcagct ctctcagtct ctccttccgg ccatagtgga actggctgaa gatgccaagt
     2041 ggagggtccg cctggctatc attgagtaca tgccgctgct ggctggccag ctgggtgtgg
     2101 agttctttga tgaaaaactc aattctttgt gtatggcctg gcttgtggat catgtctatg
     2161 ccattcggga ggcagccacc aacaacctca tgaagttagt ccagaagttc ggtacagagt
     2221 gggcccagaa taccattgtc cctaaagtgt tggtaatggc aaatgaccct aactacttac
     2281 acagaatgac cacgctcttc tgcattaatg cactgtctga agcctgtggt aaggaaataa
     2341 ccactaagca aatgttgccc atcgtgttga aaatggcagg agaccaggta gcaaatgtcc
     2401 gcttcaatgt ggccaaatca ctgcagaaaa ttggaccgat tctagacacc aacgccttgc
     2461 agggggaagt gaagccggtg ctgcagaaat tgggtcagga cgaagacatg gatgtcaaat
     2521 attttgcaca ggaagctata agtgttcttg ccttggcagg atccgactac aaagaccatg
     2581 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2641 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2821 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2881 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2941 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3001 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3061 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3121 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3181 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3241 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3301 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3361 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3421 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3481 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3541 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3601 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3781 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3841 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3901 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3961 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4021 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4081 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4201 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4261 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4321 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4381 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4441 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4501 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4561 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4621 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4681 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4741 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4801 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4861 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4921 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4981 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5041 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5101 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5161 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     5221 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5281 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5341 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5401 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5461 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5521 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5581 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5641 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5701 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5761 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5821 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5881 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5941 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6001 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6061 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6121 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6181 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6241 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6301 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6361 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6421 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6481 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6541 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6601 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6661 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6721 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6781 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6841 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6901 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6961 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7021 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7081 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7141 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7201 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7261 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7321 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7381 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7441 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7501 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7561 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7621 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7681 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7741 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7801 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7861 atgaagaatc tgcttagg
//
