LOCUS       V041154                 9305 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041154
VERSION     V041154
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..3952
                     /label="PPP1R13B(NM_015316)"
                     /note="PPP1R13B(NM_015316)"
                     /gene="PPP1R13B"
     CDS             3992..4057
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4081..4669
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4735..4783
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4985..5413
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5427..5756
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5823..6617
                     /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    6793..6914
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6963..6979)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6987..7003
                     /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(7011..7041)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7056..7077
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7365..7953)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8124..8984)
                     /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(8985..9089)
                     /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 ccatgatgcc gatgatatta actgttttct tgagcaacaa
      721 tgaacagatt ttaacagaag ttcctataac accggaaaca acctgtcgag atgttgtaga
      781 attttgcaag gaacctggag aaggcagctg ccatttagct gaagtgtgga ggggaaatga
      841 acgtcccata ccctttgatc atatgatgta cgaacatctt cagaaatggg gtccacggag
      901 ggaagaagtg aaatttttcc ttcgacacga ggactcccca actgagaaca gtgaacaagg
      961 tggccgtcag acccaagagc aacgaactca gagaaatgta ataaatgtac ctggagaaaa
     1021 acgtactgaa aatggggttg ggaatccacg tgttgaactt accctctcag agctccaaga
     1081 tatggcagct aggcaacagc agcagattga aaatcagcag cagatgttgg ttgccaagga
     1141 acagcgttta cattttctaa agcaacagga gcgccgtcag cagcagtcta tttctgaaaa
     1201 tgaaaagctt cagaaattga aagaacgagt tgaagcccag gagaacaagc tgaagaaaat
     1261 tcgtgcaatg agaggacaag tcgactacag caaaatcatg aacggcaatc tgtctgctga
     1321 aatagaaagg ttcagtgcca tgttccagga aaagaagcag gaagtacaga ctgcaatttt
     1381 aagggttgat cagcttagtc agcaattgga agatttaaag aaaggaaaac tgaatgggtt
     1441 ccagtcttac aatggcaaat tgacgggacc agcggcggtg gagttaaaaa gactgtacca
     1501 agaactacag attcgtaacc aacttaacca ggaacaaaat tcaaaacttc agcagcagaa
     1561 ggaactctta aataagcgca acatggaggt ggccatgatg gacaagcgaa tcagtgaact
     1621 gcgtgaacgt ctctatggga aaaaaattca gctgaaccgt gtgaatggca cgtcatcacc
     1681 acagtcccct ctgagcacat cgggcagggt cgctgctgtg gggccttata tccaggttcc
     1741 cagtgccgga agctttcctg tgctggggga ccctataaag ccccagtctc tcagtattgc
     1801 ctcaaatgct gctcatggaa gatccaaatc cgctaatgat ggaaactggc caacattaaa
     1861 acagaattct agctcttccg tgaaaccagt gcaggtggcc ggtgcagact ggaaggatcc
     1921 gagcgtggag gggtctgtca agcagggcac tgtctccagc cagcctgtgc ccttctcagc
     1981 actgggaccc acggagaagc cgggcatcga gattggtaaa gtgccacctc ccatcccggg
     2041 tgtaggcaag cagctgcctc caagctatgg gacataccca agtcctacac ctctgggtcc
     2101 tgggtcgaca agctccctgg aaaggaggaa ggaaggcagc ttgcccaggc ccagtgcagg
     2161 cctgccaagt cgacagaggc ccaccctgct gcccgccaca ggcagcaccc cccagccagg
     2221 ctcctcacaa cagattcagc agaggatttc cgtaccgcca agtcccacgt acccgccagc
     2281 gggaccacct gcatttccag ctggggacag caagcctgaa ctcccactga cagtggccat
     2341 taggcctttc ctggctgata aagggtcaag gccacagtct cccaggaaag gaccccagac
     2401 agtgaattca agttccatat actccatgta cctccagcaa gccacaccac ctaagaatta
     2461 ccagccggca gcacacagcg ccttaaataa gtcagttaaa gcagtgtatg gtaagcccgt
     2521 tttaccttcg ggttcaacct ctccatcgcc gctgccgttt cttcacgggt cactgtccac
     2581 gggcacacca cagcctcagc caccttcaga aagtactgag aaagagcctg agcaggatgg
     2641 ccccgccgcc cccgcagatg gcagcaccgt ggagagcctg ccacggccac tcagccccac
     2701 caagctcacg cccatcgtgc attcgccact gcgctaccag agtgatgcag acctggaggc
     2761 cctccgcagg aagctggcca acgcgccccg gcccctgaaa aagcgcagct ccatcacaga
     2821 gcccgagggc cccggcgggc ccaacatcca gaagctgctg taccagcgct tcaacaccct
     2881 ggccggtggc atggagggca cccctttcta ccagcccagc ccctcccagg acttcatggg
     2941 caccttggcc gatgtggaca atggaaacac caatgccaat ggaaacctgg aagagctccc
     3001 ccctgcccag cccacagccc cactccccgc tgagcctgcc ccgtcatcag atgccaatga
     3061 taatgagtta ccttcccccg aaccagagga gctcatctgt ccccaaacca cccaccaaac
     3121 tgccgagccg gcagaggaca ataacaacaa cgtggccacg gtccccacca cggagcagat
     3181 cccgagtcct gtggctgagg ccccatctcc aggggaagag caggtccctc cagcacctct
     3241 tccccctgcc agccaccctc ctgccacctc cacgaacaag cggaccaact tgaagaagcc
     3301 caactcggag cggacggggc acgggctgag agtccggttt aaccccctgg cactgctcct
     3361 agacgcgtct ctggaaggag agttcgatct ggtgcagagg atcatctatg aggtggaaga
     3421 tcccagcaag cccaacgacg aagggatcac cccactgcac aacgccgtct gcgccggcca
     3481 ccatcacatc gtgaagttcc tgctggattt tggtgtcaac gtgaatgctg ctgatagtga
     3541 tggatggacg ccgctgcact gcgctgcctc ttgtaacagc gttcacctct gcaaacagct
     3601 ggtggagagt ggtgccgcca tttttgcctc aaccataagc gacattgaaa ctgctgcaga
     3661 caagtgtgag gagatggagg aaggctacat ccagtgctcc cagtttctat atggggtgca
     3721 ggaaaagctg ggtgtgatga acaaaggtgt ggcgtatgct ctgtgggact acgaggccca
     3781 gaacagtgac gagctgtcct tccacgaagg ggacgccctc accatcctga ggcgcaagga
     3841 cgaaagcgag actgagtggt ggtgggctcg ccttggagac cgggagggct atgtgcccaa
     3901 aaacctgctg gggctgtatc cacggatcaa accccgacag cgaacactcg ccgaattctg
     3961 cagatatcca gcacagtggc ggccgctcga ggactacaaa gaccatgacg gtgattataa
     4021 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     4081 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4141 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4201 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4261 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4321 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4381 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4441 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4501 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4561 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4621 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     4681 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4741 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4801 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4861 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4921 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     4981 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     5161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5341 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5401 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5761 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5821 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5881 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5941 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6001 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6061 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6121 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6181 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6241 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6301 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6361 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6421 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6481 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6541 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6601 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     6661 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6721 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6781 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6841 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6901 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6961 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7021 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7141 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7201 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7261 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     7561 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7801 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7861 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7921 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7981 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8041 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     8101 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8161 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8221 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8281 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8341 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8401 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8461 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8521 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8581 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8641 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8701 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8761 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8821 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8881 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8941 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     9001 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     9061 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     9121 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     9181 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     9241 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     9301 ttagg
//
