LOCUS       pEnCMV-Piwil2(mouse)    9131 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019153
VERSION     V019153
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..3808
                     /label="Piwil2(NM_001364321)"
                     /note="Piwil2(NM_001364321)"
                     /gene="Piwil2"
     CDS             3818..3883
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3907..4495
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4561..4609
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4811..5239
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5253..5582
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5649..6443
                     /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    6619..6740
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6789..6805)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6813..6829
                     /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(6837..6867)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6882..6903
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7191..7779)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7950..8810)
                     /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(8811..8915)
                     /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 ccatgagttt gaggctagcc tgtgctacag agtcaggtcc
      721 aggtaaatct aaaactcacc cctctggtgc tttttgctcc tcttcactgt ttcccttctc
      781 tccacagcat ttgtcatcct ttcatttctt atctgagatc ccactgatag tctatgacca
      841 cagagtacag ataaggaatt ggaacagaac caacagtcgt ttctacagcc tctcaatgga
      901 tcctgtcagg ccgttgttca gggggcccac cccagtccac ccatctcagt gtgtgcggat
      961 gccaggctgt tggcctcaag ctcctagacc tttggaacca gcttggggta gggcaggacc
     1021 tgcaggcaga ggccttgtgt ttagaaaacc agaagactcc agcccaccac tccagccagt
     1081 acaaaaggat tctgtaggtt tggtgtccat gttccgtggc atgggtcttg acacagcatt
     1141 ccggcctcct tcaaaacgag aagtgcctcc tttaggcaga ggagttctag gtcgaggctt
     1201 gtctgctaac atggtccgca aggacagaga agaaccccgt tcctctttgc ctgatccttc
     1261 ggtgctggca gctggggaca gcaaactggc agaggcttct gttggttgga gtagaatgct
     1321 gggaagaggt agttctgagg tctctctgtt accactggga cgagcagcca gtagtatagg
     1381 cagaggaatg gacaaacctc ccagtgcctt tggcctcaca gctcgggatc ccccacggct
     1441 gccacagcct ccagctctgt ctccaacttc actgcactct gccgatcccc ctccggtcct
     1501 gactatggaa cgaaaggaaa aagagctttt ggtcaagcaa ggatcaaaag gaactcctca
     1561 gtctttggga ctgaacctca tcaaaatcca gtgtcataac gaagcagttt atcagtacca
     1621 tgtgactttc agccccagtg tggaatgcaa aagcatgagg tttggcatgt tgaaggacca
     1681 ccagtctgtc actggaaacg tcactgcttt tgatggctct attctttatc ttcctgttaa
     1741 gcttcaacaa gttgttgagt taaaaagtca gaggaaaact gacgatgccg agatcagtat
     1801 caagattcag ctgacaaaga tcctggagcc gtgttctgac ctgtgcatcc ccttctacaa
     1861 tgttgtcttc cggcgggtaa tgaaacttct ggatatgaag cttgtgggga gaaacttcta
     1921 tgaccctaca agtgccatgg tactgcagca acacagattg cagatctggc ctggctatgc
     1981 ggctagtatc cggaggacag acgggggtct cttcctgctc gctgatgtct ctcataaggt
     2041 cattcggaac gactctgtgc tggatgtcat gcatgctatc taccagcaga acaaggagca
     2101 cttccaggac gagtgcagca agcttctggt tggcagcatt gtcatcacgc gctacaacaa
     2161 tcgtacctac cgaatcgatg atgtggactg gaacaagacc cctaaagaca gctttgtcat
     2221 gtcggacggg aaggaaatca cattcctgga atactacagc aaaaactatg ggatcacagt
     2281 caaggaagat gaccagccgc tgctgatcca ccggcccagt gagagacaga ataaccatgg
     2341 catgttgctg aagggcgaga tcctgctgct gcccgagctc tccttcatga cggggatccc
     2401 tgagaagatg aagaaggact tcagggccat gaaggacttg actcagcaga ttaacctgag
     2461 ccccaagcag caccacggtg ctttggaatg cctgctgcag agaatttcac aaaacgagac
     2521 agccagcaat gagctgaccc gctgggggct cagtctgcat aaagatgtcc acaagattga
     2581 aggtcggctt ctgccaatgg agaggatcaa cttaaggaac acttcatttg tcacatcgga
     2641 ggacctgaac tgggttaagg aagtgaccag agatgcttcc attctaacta ttcccatgca
     2701 tttctgggca ctcttttatc caaagagagc aatggaccaa gccagagaac tggttaacat
     2761 gttggaaaag attgccgggc ccattggcat gcgcataagc cccccagcct gggttgagct
     2821 gaaggatgac cgaatagaga cctatatcag gaccattcag tccttactgg gagttgaggg
     2881 gaagatacaa atggtcgttt gcatcatcat gggcacacgt gatgatctct atggagccat
     2941 caagaagctg tgctgcgtgc agtccccagt gccctcacag gtcatcaatg tccgaaccat
     3001 tggtcagccc accaggcttc ggagcgtggc tcagaaaatt ttacttcaga tgaactgtaa
     3061 actgggtggt gagctctggg gagtggatat tccgctgaaa caactaatgg tgattggaat
     3121 ggatgtgtac catgacccca gcagaggcat gcgctctgtg gtcggcttcg tggccagcat
     3181 aaatctcaca ctcaccaaat ggtactcgag ggtggtgttc cagatgccac atcaggagat
     3241 tgtggacagc ctgaagctct gcctggtggg ttccttgaaa aagtattatg aggtgaacca
     3301 ttgtctccca gagaaaattg tggtgtaccg agatggagtg tctgatggcc agctaaagac
     3361 agttgccaac tacgagatcc ctcagctgca gaagtgtttt gaagcctttg ataactacca
     3421 ccccaagatg gtggtgtttg tagttcagaa gaaaatcagc accaatctgt accttgctgc
     3481 tcctgatcac ttcgtaaccc cctcccccgg gactgtggtt gatcatacca taaccagctg
     3541 tgagtgggtg gatttctacc ttcttgccca tcatgtgcga cagggctgtg gcatacctac
     3601 acactacatc tgtgttctga acactgcaaa tctgagccct gatcacatgc agaggttgac
     3661 tttcaaacta tgccacatgt actggaattg gcctggtacc atccgagttc cagctccttg
     3721 caagtatgcc cacaagctag ctttcctgtc cggacagatt ttgcatcatg agccagccat
     3781 ccagctgtgt gggaacctgt tcttcctgta actcgaggac tacaaagacc atgacggtga
     3841 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat
     3901 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3961 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4021 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4081 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4141 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4201 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4261 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4321 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4381 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4441 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     4501 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4561 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     4621 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4681 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4741 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     4801 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4861 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4921 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     4981 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5041 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5101 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5161 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     5221 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5281 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5341 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5401 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5461 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5521 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5581 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     5641 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5701 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5761 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5821 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5881 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5941 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6001 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6061 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6121 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6181 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6241 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6301 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     6361 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6421 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     6481 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6541 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6601 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6661 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6721 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6781 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6841 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6901 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7381 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7621 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8941 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     9001 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9061 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9121 atctgcttag g
//
