LOCUS       V042848                 7616 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042848
VERSION     V042848
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             890..934
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             941..1054
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             1061..1087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1094..3136
                     /label="PAK6(NM_020168)"
                     /note="PAK6(NM_020168)"
                     /gene="PAK6"
     polyA_signal    3183..3407
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3453..3881
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3895..4224
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4291..5085
                     /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    5259..5380
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5429..5445)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5453..5469
                     /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(5477..5507)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5522..5543
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5831..6416)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6587..7447)
                     /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(7448..7552)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttaccatga aagaaaccgc
      901 tgctgctaaa ttcgaacgcc agcacatgga cagcggtgga atggacgaga agaccaccgg
      961 ctggagaggc ggccacgtgg tggagggcct ggccggcgag ctggagcagc tgagagccag
     1021 actggagcac caccctcagg gccagagaga acctggtacc tacccatacg atgttccaga
     1081 ttacgctgga tccatgttcc gcaagaaaaa gaagaaacgc cctgagatct cagcgccaca
     1141 gaacttccag caccgtgtcc acacctcctt cgaccccaaa gaaggcaagt ttgtgggcct
     1201 ccccccacaa tggcagaaca tcctggacac actgcggcgc cccaagcccg tggtggaccc
     1261 ttcgcgaatc acacgggtgc agctccagcc catgaagaca gtggtgcggg gcagcgcgat
     1321 gcctgtggat ggctacatct cggggctgct caacgacatc cagaagttgt cagtcatcag
     1381 ctccaacacc ctgcgtggcc gcagccccac cagccggcgg cgggcacagt ccctggggct
     1441 gctgggggat gagcactggg ccaccgaccc agacatgtac ctccagagcc cccagtctga
     1501 gcgcactgac ccccacggcc tctacctcag ctgcaacggg ggcacaccag caggccacaa
     1561 gcagatgccg tggcccgagc cacagagccc acgggtcctg cccaatgggc tggctgcaaa
     1621 ggcacagtcc ctgggccccg ccgagtttca gggtgcctcg cagcgctgtc tgcagctggg
     1681 tgcctgcctg cagagctccc caccaggagc ctcgcccccc acgggcacca ataggcatgg
     1741 aatgaaggct gccaagcatg gctctgagga ggcccggcca cagtcctgcc tggtgggctc
     1801 agccacaggc aggccaggtg gggaaggcag ccctagccct aagacccggg agagcagcct
     1861 gaagcgcagg ctattccgaa gcatgttcct gtccactgct gccacagccc ctccaagcag
     1921 cagcaagcca ggccctccac cacagagcaa gcccaactcc tctttccgac cgccgcagaa
     1981 agacaacccc ccaagcctgg tggccaaggc ccagtccttg ccctcggacc agccggtggg
     2041 gaccttcagc cctctgacca cttcggatac cagcagcccc cagaagtccc tccgcacagc
     2101 cccggccaca ggccagcttc caggccggtc ttccccagcg ggatcccccc gcacctggca
     2161 cgcccagatc agcaccagca acctgtacct gccccaggac cccacggttg ccaagggtgc
     2221 cctggctggt gaggacacag gtgttgtgac acatgagcag ttcaaggctg cgctcaggat
     2281 ggtggtggac cagggtgacc cccggctgct gctggacagc tacgtgaaga ttggcgaggg
     2341 ctccaccggc atcgtctgct tggcccggga gaagcactcg ggccgccagg tggccgtcaa
     2401 gatgatggac ctcaggaagc agcagcgcag ggagctgctc ttcaacgagg tggtgatcat
     2461 gcgggactac cagcacttca acgtggtgga gatgtacaag agctacctgg tgggcgagga
     2521 gctgtgggtg ctcatggagt tcctgcaggg aggagccctc acagacatcg tctcccaagt
     2581 caggctgaat gaggagcaga ttgccactgt gtgtgaggct gtgctgcagg ccctggccta
     2641 cctgcatgct cagggtgtca tccaccggga catcaagagt gactccatcc tgctgaccct
     2701 cgatggcagg gtgaagctct cggacttcgg attctgtgct cagatcagca aagacgtccc
     2761 taagaggaag tccctggtgg gaacccccta ctggatggct cctgaagtga tctccaggtc
     2821 tttgtatgcc actgaggtgg atatctggtc tctgggcatc atggtgattg agatggtaga
     2881 tggggagcca ccgtacttca gtgactcccc agtgcaagcc atgaagaggc tccgggacag
     2941 ccccccaccc aagctgaaaa actctcacaa ggtctcccca gtgctgcgag acttcctgga
     3001 gcggatgctg gtgcgggacc cccaagagag agccacagcc caggagctcc tagaccaccc
     3061 cttcctgctg cagacagggc tacctgagtg cctggtgccc ctgatccagc tctaccgaaa
     3121 gcagacctcc acctgctgac tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3181 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3241 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3301 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3361 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3421 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3661 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3721 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3781 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3841 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3901 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3961 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4081 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4141 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4201 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4261 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4321 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4381 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4441 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4501 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4561 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4621 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4681 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4741 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4801 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4861 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4921 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4981 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5041 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5101 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5161 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5221 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5401 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5461 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5521 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5581 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5641 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5701 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5761 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5821 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5881 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5941 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6001 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6061 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6121 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6181 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6241 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6301 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6361 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6421 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6481 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6541 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6601 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6661 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6721 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6781 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6841 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6901 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6961 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7021 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7081 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7141 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7201 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7261 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7321 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7381 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7441 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7501 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7561 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
