LOCUS       V022654                 7805 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022654
VERSION     V022654
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"
     CDS             683..2485
                     /label="PPP2R1B(NM_002716)"
                     /note="PPP2R1B(NM_002716)"
                     /gene="PPP2R1B"
     CDS             2492..2557
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2581..3169
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3235..3283
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3485..3913
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3927..4256
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4323..5117
                     /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    5293..5414
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5463..5479)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5487..5503
                     /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(5511..5541)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5556..5577
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5865..6453)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6624..7484)
                     /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(7485..7589)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcggg cgcatcagag ctcgggaccg gcccaggagc
      721 agcgggtgga gatggagatg attcgctata cccgatcgcg gttttaatcg acgagctccg
      781 caatgaagac gtgcagctcc gactcaacag tattaagaag ttatcaacaa ttgccctagc
      841 acttggagta gaaaggaccc gaagtgaatt gttgccattt cttacagata caatttatga
      901 tgaagatgag gtactattag ctcttgctga gcagctggga aatttcactg gcctagtggg
      961 aggtcctgac tttgcccact gtctgctgcc tcctttggaa aatctggcaa ctgtggaaga
     1021 gactgttgtt cgtgacaagg ctgtggagtc cctgagacag atctcccagg agcatactcc
     1081 tgttgctctg gaagcttatt ttgtacctct ggtgaaacgc ttagcaagtg gggattggtt
     1141 cacctctcgc acatctgcat gtggtttgtt cagcgtttgc tatcccaggg catcaaatgc
     1201 tgttaaagca gaaatcagac agcaattccg ttccttgtgc tcagatgaca caccaatggt
     1261 acgacgtgct gctgcttcca aattgggtga atttgcaaaa gttttggaat tagacagtgt
     1321 gaaaagtgaa attgttccac tgttcactag tctagcttca gatgaacagg attcagtgcg
     1381 cctccttgct gtggaagctt gtgtcagtat tgcccagtta ttgtctcagg atgaccttga
     1441 gactttggtg atgcctacac ttcgacaagc agcagaagat aaatcttggc gcgttcgcta
     1501 tatggtggct gacagatttt cagagctcca gaaagccatg ggtcctaaaa tcaccctaaa
     1561 tgacctcatc cccgcctttc agaacctact taaagactgt gaagctgaag tccgggcagc
     1621 tgctgcccac aaagtaaaag aacttggtga gaacttgccc attgaagata gagagaccat
     1681 aattatgaat caaattctgc cttatataaa ggaattagta tccgatacca atcaacatgt
     1741 caaatcggct ctagcttctg taattatggg attgtctact attttgggca aagaaaatac
     1801 cattgaacat cttctacctc ttttcttagc tcagttaaag gatgagtgtc ctgacgttcg
     1861 tttgaatatc atctccaatt tggattgtgt aaatgaagtg attggaatcc gtcagctctc
     1921 tcagtctctc cttcctgcca tagtggagct ggcagaagat gccaaatgga gggtccgcct
     1981 ggccatcatt gagtatatgc cgctgctggc aggccagctg ggtgtggaat tctttgatga
     2041 aaagctgaat tctttatgta tggcttggct cgtggaccat gtatacgcca tccgagaagc
     2101 tgccaccaac aacctcatga aactagttca gaagtttggt acagagtggg cccaaaatac
     2161 tattgttccc aaagtgttag taatggcaaa tgatcctaat tacttgcata gaatgaccac
     2221 tttattctgc attaatgcac tgtctgaggc ctgtggtcag gaaataacta ctaagcaaat
     2281 gctgcccatc gtattaaaaa tggcaggaga ccaagtagca aatgttcgct tcaatgtggc
     2341 caaatctcta caaaagattg gaccaattct agataccaat gctttacagg gagaagtgaa
     2401 gccagtacta cagaagttag gtcaagatga agacatggat gtcaaatact ttgcacagga
     2461 agctataagt gttcttgcat tggcactcga ggactacaaa gaccatgacg gtgattataa
     2521 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     2581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2941 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3001 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3121 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     3181 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3241 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3301 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3361 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3421 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     3481 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3541 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3601 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3661 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3721 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3781 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3841 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3901 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4021 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4081 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4141 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4201 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4261 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4321 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4501 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5101 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     5161 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5221 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5281 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5341 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5401 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5461 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5521 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5701 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5761 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5821 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5881 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5941 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6001 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     6061 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6121 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6181 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6241 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6301 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6361 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6481 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7621 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     7681 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7741 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7801 ttagg
//
