LOCUS       V033854                 8632 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033854
VERSION     V033854
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             706..3291
                     /label="Piwil1(NM_021311)"
                     /note="Piwil1(NM_021311)"
                     /gene="Piwil1"
     CDS             3298..3324
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3328..3354
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3358..3384
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3408..3996
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4062..4110
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4312..4740
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4754..5083
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5150..5944
                     /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    6120..6241
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6290..6306)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6314..6330
                     /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(6338..6368)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6383..6404
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6692..7280)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7451..8311)
                     /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(8312..8416)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgac tggccgagcc
      721 cgagctcggg cccgcggcag ggcacgaggt caggagacgg tgcagcatgt tggggctgct
      781 gcgagccagc aacctgggta catcccaccg agacctcaac agtcccccac agagggggac
      841 ttggttggcc gaggacgaca gagggggatg gtagtcggag ccacatccaa gtcacaagaa
      901 ctgcagatct cagctgggtt tcaggagctg tcactggcag agagaggagg gcgtcgccga
      961 gacttccatg accttggtgt gaacaccaga cagaaccttg accatgtcaa agagtcaaag
     1021 acaggctcct ctggcatcat tgtgaagctg agcaccaacc acttccggct gacctcgcgc
     1081 ccacagtggg ccctgtatca gtaccacatc gactacaatc ccctgatgga ggcccgaagg
     1141 cttcgctccg cactgctctt ccagcatgaa gacctcattg gaaggtgtca tgctttcgat
     1201 gggacaatat tgtttttacc taagagacta cagcacaagg tcacagaagt attcagtcag
     1261 actcggaatg gggaacacgt gaggatcacc atcaccctga ccaacgagct gccgcccacc
     1321 tcgcccacct gcctgcagtt ctataacatc atcttcagga ggctcttgaa aatcatgaat
     1381 ttgcaacaaa ttggacggaa ttattacaat ccaagtgacc cgattgatat tccaaaccac
     1441 aggttggtga tctggcccgg cttcaccacc tccatccttc agtatgagaa caacatcatg
     1501 ctctgcacag acgtcagcca caaggtgctc cgcagcgaga ctgtcctaga cttcatgttc
     1561 aatctatacc agcagacaga ggagcacaag ttccaggagc aagtgtcgaa ggagctcata
     1621 ggcctcatcg ttctcaccaa gtacaataac aagacctacc gggtggatga cattgactgg
     1681 gaccagaatc caaagagcac cttcaagaag gcggatggct cggaggtcag cttcctggag
     1741 tactacagga agcaatacaa ccaggagatc acggacctga agcagccggt gctggtgagc
     1801 caacccaagc ggaggagagg ccccggcggc accctgcctg gcccagctat gctcatccct
     1861 gaactctgct atctcacagg cctgactgat aaaatgcgca atgatttcaa tgtgatgaag
     1921 gacctggcag tgcacacgcg gctgacccct gagcagcggc agcgggaggt gggccgcctc
     1981 atcgactaca tccacaagga tgacaatgtg cagagagagc ttcgagactg gggcctgagc
     2041 ttcgactcaa acttgctgtc cttctctgga agaatcttac aatctgagaa gatccaccag
     2101 ggcggaaaga cgtttgatta caacccacaa tttgcagact ggtccaaaga aacaagaggc
     2161 gcgccgctga tcagcgtgaa gccattggat aactggctgc tgatctatac ccgcaggaat
     2221 tatgaagcag ccaactcact gatacagaac ctgttcaaag tgactccagc catgggcatc
     2281 cagatgaaaa aggcaatcat gatcgaggtg gatgacagaa cagaagctta tctgagagcc
     2341 ttgcagcaga aggtgacgtc agacactcag atagttgtct gtctcttgtc aagtaatcgg
     2401 aaggacaaat atgatgccat caagaagtac ttgtgtacag actgccccac cccaagtcag
     2461 tgtgtggtgg cccggaccct gggcaagcag caaacagtca tggccattgc caccaagatc
     2521 gccctgcaga tgaactgcaa gatgggaggc gagctctggc gggtggacat gcccctgaaa
     2581 ctggcaatga tcgtgggcat cgactgttac catgacacca cagctgggcg gaggtccatc
     2641 gcaggattcg tcgccagcat caatgaaggg atgacccgct ggttctcccg ctgcgtcttt
     2701 caggaccgcg ggcaggagct ggtggatggt ctcaaggtgt gcttgcaagc tgctctgagg
     2761 gcttggagtg gctgcaatga atacatgccc agccgtgtca tcgtgtaccg agacggtgtg
     2821 ggggacgggc agctgaagac cctggtcaat tatgaggtcc cacagttcct agattgcctc
     2881 aagtcagtcg ggagaggtta caacccaaga ctgactgtaa tcgtggtgaa gaagcgtgtc
     2941 aatgccaggt tttttgctca gtctggggga agacttcaga accctcttcc agggacagtc
     3001 atcgatgtgg aagtcaccag accagagtgg tatgactttt tcatcgtgag ccaggcagtg
     3061 agaagcggga gtgtgtcccc aacacactac aatgtcatct atgacagcag tggcctgaag
     3121 cccgaccaca tccagcggct gacatacaag ctctgccacg tgtactataa ttggcctgga
     3181 gtcatccgag tccctgcacc ttgccagtat gcacacaagc tggccttcct cgtgggccag
     3241 agcatccaca gagagccaaa cctctccctg tccaaccgcc tctactacct cctcgagtac
     3301 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat
     3361 ccttatgatg tcccagacta cgcatagtga gggcccgata tctcgacaat caacctctgg
     3421 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3481 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3541 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3601 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3661 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3721 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3781 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     3841 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3901 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3961 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga
     4021 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4081 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4141 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4201 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4261 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
     4321 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4381 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4441 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     4501 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4561 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4621 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4681 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4741 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4801 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4921 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4981 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     5041 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     5101 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5161 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5221 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5281 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5341 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5401 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5461 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5521 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5581 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5641 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5701 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5761 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5821 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5881 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5941 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6001 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6061 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     6121 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6181 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6241 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6301 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6361 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6421 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6481 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6541 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6601 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6661 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6721 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6781 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6841 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     6901 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6961 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7021 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7081 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7141 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7201 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7261 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7321 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7381 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7441 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7501 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7561 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7621 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7681 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7741 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7801 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7861 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7921 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7981 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8041 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8101 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8161 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8221 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8281 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8341 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8401 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8461 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag
     8521 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     8581 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
