LOCUS       V034089                 6373 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034089
VERSION     V034089
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..1863
                     /label="Arpc2(NM_001106919)"
                     /note="Arpc2(NM_001106919)"
                     /gene="Arpc2"
     CDS             1870..1935
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1973..2197
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2243..2671
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2685..3014
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3081..3875
                     /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    4049..4170
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4219..4235)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4243..4259
                     /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(4267..4297)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4312..4333
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4621..5206)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5377..6237)
                     /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(6238..6342)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgatcc tgctggaggt gaacaaccgc atcatcgagg agacgctcgc gctcaagttc
     1021 gagaacgcgg ccgccggaaa caaaccagaa gctgtagaag taacatttgc agattttgat
     1081 ggcgtccttt atcatatttc aaatcctaat ggagacaaaa caaaagtgat ggtcagtatt
     1141 tctttgaaat tctacaagga acttcaggca cacggagctg atgagttact aaagagagta
     1201 tacggaagtt tcttggtgaa cccagaatca ggatacaatg tgtctttgct atatgacctg
     1261 gaaaatctac ctgcatccaa ggattctatt gtgcatcagg ctggcatgtt gaagcgaaat
     1321 tgttttgcct ctgtgtttga gaaatacttc caattccaag aagagggcaa agaaggagag
     1381 aacagagcag ttatccatta tagagatgat gagaccatgt atgtggaatc taaaaaagac
     1441 agagtcacag tagtcttcag cacagttttt aaggatgacg acgatgtggt cattggaaag
     1501 gtgttcatgc aggagttcaa agaaggacga agagccagcc acacagcccc ccaggtcctt
     1561 tttagccaca gggaacctcc tctagaactg aaagacacag atgctgccgt gggtgacaac
     1621 attggctaca tcacctttgt gctgttccct cgccacacca atgccactgc tcgagacaac
     1681 accatcaacc tgatccacac gttccgggac tatctgcact accacattaa gtgctctaag
     1741 gcctatattc atacacgaat gcgagcaaaa acatccgact tccttaaggt gctcaaccgt
     1801 gcacgcccag atgccgagaa aaaagaaatg aaaacaatca cggggaagac tttttcatcc
     1861 cgcctcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     1921 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1981 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2041 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2101 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2161 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2221 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2281 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2341 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2401 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2461 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2521 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2581 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2641 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2701 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2761 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2821 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2881 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2941 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3001 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3061 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3121 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3181 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3241 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3301 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3361 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3421 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3481 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3541 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3601 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3661 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3721 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3781 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3841 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3901 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3961 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4021 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4081 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4141 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4201 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5041 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     5161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6361 gccacctgac gtc
//
