LOCUS       V027539                 8369 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027539
VERSION     V027539
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             941..3859
                     /label="PIWIL2(NM_018068)"
                     /note="PIWIL2(NM_018068)"
                     /gene="PIWIL2"
     CDS             3866..3931
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3969..4193
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4239..4667
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4681..5010
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5077..5871
                     /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    6045..6166
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6215..6231)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6239..6255
                     /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(6263..6293)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6308..6329
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6617..7202)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7373..8233)
                     /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(8234..8338)
                     /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 atccgccacc atggatcctt tccgaccatc
      961 gttcaggggc cagtctccta tccacccatc ccagtgccag gctgtacgga tgccaggctg
     1021 ttggccacaa gcttctaaac ctttggaccc agctctgggc aggggagcac ctgcaggcag
     1081 aggccatgta tttggaaagc cagaggaacc aagcacacag agggggccag cacaaaggga
     1141 gtctgtgggt ttggtctcca tgttccgagg cctgggcatt gaaacagttt ctaagacccc
     1201 tctgaaacgg gaaatgcttc catcaggtag aggcatttta ggtcgaggct tgtctgctaa
     1261 tctggtacgc aaggacaggg aggaactctc tcccactttt tgggatccaa aagtgttggc
     1321 ggctggggac agcaagatgg cagagacctc cgttggttgg agtaggacgc ttggaagagg
     1381 gagttcagat gcgtctttat taccactggg aagagcagca ggtggtatca gcagagaagt
     1441 ggacaagcct ccctgtacct tcagcacacc gtcccggggt cccccgcagc tgtcatcacc
     1501 accagctctg ccccagtctc ccctgcactc tccagatcgc cctctggtcc tgactgtgga
     1561 acacaaggaa aaagagctta ttgtgaagca aggatcaaaa ggaacacctc agtctttggg
     1621 actgaacctc gtcaaaatac agtgtcataa tgaagcagtt tatcaatatc atgtgacttt
     1681 cagccccaat gtggagtgca aaagcatgag gttcggcatg ttgaaggacc atcaagctgt
     1741 caccggcaac gtcactgcgt ttgatggatc tattctctat ctgcctgtta agcttcaaca
     1801 agttcttgag ttaaaaagtc aaaggaaaac agacagtgct gaaatcagca ttaagattca
     1861 gatgacaaag atcctggagc cctgctctga cctgtgcatt cccttctaca atgttgtttt
     1921 ccgtcgggta atgaaacttt tagatatgaa gcttgtgggg agaaactttt atgaccctac
     1981 aagtgctatg gtactacagc aacacagatt gcagatctgg ccaggctatg cagctagcat
     2041 ccgaaggaca gatggagggc tcttcctgct agctgatgtc tcccataagg tcattcggaa
     2101 tgactgtgtg ctggatgtca tgcatgccat ttatcagcag aataaagaac acttccagga
     2161 tgagtgtact aagcttctgg ttggcaatat tgttatcacc cgatataaca atcgtaccta
     2221 tcgtattgat gatgtggatt ggaataagac tccaaaggat agcttcacga tgtctgatgg
     2281 gaaagagatc acattcttgg aatactacag caaaaattat gggatcacag ttaaggaaga
     2341 ggaccagcca ttgctgattc acaggcccag tgagagacag gataatcatg ggatgctgct
     2401 aaaaggggaa atcctgctgc tgcctgagct ttcttttatg accggaatcc cagagaagat
     2461 gaagaaggac ttcagagcca tgaaggattt ggctcagcaa atcaatctga gccccaagca
     2521 acaccatagt gctttggaat gcttgctgca aagaattgca aagaacgagg cagccaccaa
     2581 tgaactgatg cgttgggggc tccgtctgca aaaggatgta cataagattg aaggacgtgt
     2641 tctgccaatg gaaagaatta acttaaaaaa tacttcgttt atcacatctc aggaactaaa
     2701 ctgggttaag gaagtaacca gagacccttc catcttgact atccccatgc atttctgggc
     2761 acttttttac ccaaagagag caatggacca ggctcgagaa ctggtcaaca tgttggagaa
     2821 gatagccggc cccattggca tgcgtatgag cccaccggcc tgggttgaac taaaggatga
     2881 ccgaatagag acttatgtca gaaccattca atccacgtta ggagctgagg ggaagataca
     2941 gatggttgtt tgcatcatca tgggcccacg tgatgatctc tatggggcca tcaagaagct
     3001 gtgctgtgtg cagtccccag tgccctccca ggttgtcaat gttcgaacca ttggtcagcc
     3061 caccaggctt cggagtgtgg cccagaagat tttacttcag attaactgta aattgggtgg
     3121 tgagctctgg ggagtggata ttcctctgaa acagttaatg gtgatcggga tggatgttta
     3181 ccatgacccc agtagaggca tgcgctccgt ggttggcttc gtggcaagca tcaatctcac
     3241 cctcacaaaa tggtattccc gggtggtgtt ccagatgccg catcaggaga ttgtggacag
     3301 cctgaagcta tgcctcgtgg gctccttaaa aaagttttat gaggtgaacc actgtctacc
     3361 agagaagatt gtggtgtacc gtgatggagt gtctgatggc caactgaaga cagttgccaa
     3421 ctatgagatt cctcaactac agaagtgttt tgaagctttt gagaattatc agcccaagat
     3481 ggtggtgttt gtagttcaga agaaaatcag tactaatcta tatctggctg ctcctcagaa
     3541 ctttgtaact cccactcctg gaactgtggt agatcataca ataacaagct gtgagtgggt
     3601 ggatttctat cttcttgccc atcatgtacg gcagggctgt ggcattccta cgcattatgt
     3661 ctgtgttctc aacaccgcaa acctgagccc tgatcatatg cagaggctga ctttcaaact
     3721 gtgccacatg tactggaatt ggcctggcac catcagagtt ccagctcctt gcaagtatgc
     3781 ccacaagcta gctttcctgt caggacacat cttgcatcat gaaccagcca tccagctgtg
     3841 cgagaacctg ttcttcctgc tcgaggacta caaagaccat gacggtgatt ataaagatca
     3901 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     3961 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4021 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4081 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4141 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4201 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4261 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4321 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4381 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4441 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4501 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4561 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4621 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4681 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4741 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4801 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4861 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4921 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4981 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5041 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5101 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5161 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5221 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5281 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5341 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5401 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5461 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5521 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5581 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5641 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5701 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5761 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5821 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5881 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5941 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6001 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6181 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6241 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6301 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6361 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6421 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6481 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6541 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6601 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6661 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6721 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6781 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6841 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6901 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6961 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7021 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7081 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7141 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctgacgtc
//
