LOCUS       V025586                 8640 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025586
VERSION     V025586
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3320
                     /label="Trim71(NM_001042503)"
                     /note="Trim71(NM_001042503)"
                     /gene="Trim71"
     CDS             3327..3392
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3416..4004
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4070..4118
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4320..4748
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4762..5091
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5158..5952
                     /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    6128..6249
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6298..6314)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6322..6338
                     /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(6346..6376)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6391..6412
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6700..7288)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7459..8319)
                     /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(8320..8424)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc ttcgttcccg gagaccgact
      781 tccagatctg cctgctgtgc aaggagatgt gcggctcgcc cgcgccgctc tcctccaact
      841 cgtccgcctc gtcgtcgtcc tcgcagactt ccacgtcgtc ggcgggcggc ggcggccccg
      901 gggcggcggc gcggcgtctt cacgtcttgc cctgcctgca cgccttctgc cggccgtgcc
      961 tcgaggcgca ccggctgccc gctcccggag gcgcgggccc cgccgaggcg ctgaagctgc
     1021 gctgccccgt atgtgaccag aaggtggtcc tggccgaggc ggcgggcatg gacgcgctgc
     1081 catcctccgc cttcctgctc agcaacctgc tggacgccgt ggtggccacc gccgaagagc
     1141 cgccgcccaa gaacgggcgc gcgggcggcg gccccggggg cgcgggcggc cacagcaacc
     1201 accggcacca tgcgcaccac ccggcccagc gcgccgccgc tccagcgccg cagccgccgc
     1261 ccggcccggc cgcctccccc gggtcgctgc tgatgcgccg gccgcacggc tgcagctcct
     1321 gcgacgaggg caacgccgcc tcgtcgcgct gcctcgactg ccaggaacac ctgtgcgaca
     1381 actgtgtccg cgcgcaccag cgcgtccgcc tcaccaagga ccattacatc gagcgcggcc
     1441 ctccggggcc tgcggccgcc tccgcggcac agcagctggg gctcgggccg cccttcgcgg
     1501 gcgcgccctt ctccattctc tcggtgttcc cggagcgcct cggcttctgc cagcaccacg
     1561 acgatgaggt cctgcatctc tactgtgaca cctgctctgt ccccatctgc agagagtgca
     1621 cgctgggccg ccacggtggc cacagcttcg cctacctcca ggatgccctg caggactctc
     1681 gggcactcac catccagctg ctggctgatg cccagcaggg ccgccaggct ctccagctga
     1741 gcatcgagca ggcccagact gtggcagaac aggtggagat gaaggcgaag gtggtccagt
     1801 ctgaggtcaa ggccgtcaca gctcgccaca agaaggccct ggaggatcgg gagtgtgagc
     1861 tgttgtggaa ggtggagaag atccggcagg tgaaggcgaa gtctctgttc ctgcaggtgg
     1921 agaagctgcg gcagagtctc agcaagctgg agagcaccat cagcgccgtg cagcaggtcc
     1981 tggaagaggg ccgcgccctg gacatcctgc tggcccgaga ccggatgctg gcccaggtgc
     2041 aggagctcaa gaccatccga ggcctcctgc agccccaaga agatgaccgc attatgttca
     2101 cgccccctga ccaggccctg tacctcgccc tcaagtctat tggctttgtc agcagcgggg
     2161 ccttcgcccc gctcacgaag gctacaggag atggcatcaa gcgtgccctt caggggaagg
     2221 tggcctcttt cactgtcatg ggctacgacc acgatggtga gccccgccat tcgggaggtg
     2281 acctgatgtc agttgtggtc ctcggccccg acggaaacct gtttggtgca gaagtgagcg
     2341 accagcagaa tggtacttac atagtgagct acaggcccca gcttgagggt gagcacttgg
     2401 tgtcggtgac attgtacaat cagcatatag agaacagccc cttcaaggtc gtggtcaagt
     2461 cgggccgcag ctacgtgggc atcgggctcc ccgggctgag cttcggcagt gaaggtgacg
     2521 gggagggcaa gctctgcagg ccctggggcg tgagtgttga taaggagggt ttcatcatag
     2581 tggctgaccg aagcaataac agaatccagg tgtttaagcc ctgtggttcc ttccaccaca
     2641 agtttggtac ccttgggtct cgtcctgggc agtttgaccg accggctggg gtggcttgtg
     2701 acgcctcccg caggatcata gtggccgaca aagacaatca tcgcattcag atcttcacct
     2761 tcgagggcca gttccttctt aagtttggtg agaaaggcac caagaacggg cagtttaact
     2821 acccgtggga tgtagcagtg aattctgagg gcaagatcct cgtttcagac acccggaatc
     2881 atcgcatcca gttgttcgga ccggacgggg tcttcctgaa caagtacggc ttcgagggat
     2941 ctctctggaa gcacttcgac tctccacggg gcgtggcttt caacaatgaa ggtcacctgg
     3001 tagtcaccga cttcaacaac caccggcttc tggttattca ccctgattgc caatctgccc
     3061 gcttcttggg ttcggagggc tcgggcaatg ggcagttcct gcgaccacag ggtgtggctg
     3121 tggaccagga agggagaatc atcgtagccg attccagaaa tcaccgcgtg cagatgtttg
     3181 aggccaatgg cagcttcctg tgcaagtttg gtgctcaagg gagtggcttc gggcagatgg
     3241 accgcccctc aggtatcgcg gtcaccccgg atggattgat cgtcgtggtg gactttggca
     3301 acaatcgaat cctcatcttc ggatccgact acaaagacca tgacggtgat tataaagatc
     3361 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     3421 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3481 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3541 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3601 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3661 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3721 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3781 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3841 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3901 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3961 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     4021 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4081 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4141 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4201 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4261 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     4321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4441 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4681 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4741 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4861 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4921 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4981 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5041 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5101 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5161 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5221 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5281 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5341 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5401 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5461 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5521 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5581 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5641 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5701 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5761 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5821 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5881 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5941 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     6001 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6061 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6121 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6181 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6241 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7141 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8461 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     8521 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     8581 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
