LOCUS       V047448                 8875 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047448
VERSION     V047448
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..4365
                     /label="Trp53bp2(NM_173378)"
                     /note="Trp53bp2(NM_173378)"
                     /gene="Trp53bp2"
     CDS             4372..4437
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4475..4699
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4745..5173
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5187..5516
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5583..6377
                     /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    6551..6672
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6721..6737)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6745..6761
                     /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(6769..6799)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6814..6835
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7123..7708)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7879..8739)
                     /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(8740..8844)
                     /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 accatgcggt tcgggtccaa aatgatgccg atgttcctga cagtttacct cagtaacagt
     1021 gagcagcact tcacagaagt tcctgttacc ccagagacaa tatgcagaga tgtggtggat
     1081 ctgtgcaagg agcctggaga gaacgactgc catttagccg aagtgtggtg tggctctgaa
     1141 cgcccagttg ctgataacga acggatgttt gatgttcttc agcggtttgg aagtcaaagg
     1201 aatgaagttc gcttctttct tcgtcatgaa cgccccccta acagggacat tgtgagtgga
     1261 ccgagatccc aagatccaag tgtaaaaaga aatggtgtca aagttcccgg cgagcatcgg
     1321 aggaaggaga atggggttaa cagtcctagg ctggacctga cgcttgctga gctccaggag
     1381 atggcatctc gccagcagca gcagatcgag gcccagcaac aaatgctggc tactaaggag
     1441 caacgcttaa agtttttaaa acagcaagat caacgtcaac agcaacaagc tgctgaacag
     1501 gagaaactta agaggcttag agaaatagct gaaagtcagg aagctaagct taagaaagtg
     1561 agagcgctaa agggccatgt ggagcaaaag aggctgagca acggaaaact cgtggaggag
     1621 atcgagcaga tgaatagctt gttccagcag aagcagaggg agctggtgct ggctgtgtcg
     1681 aaagtggagg agctgacgag gcagctggag atgctgaaga acggcaggat cgatggccac
     1741 cacgacaacc agtcggccgt ggctgagctg gaccgccttt acaaggagct gcagttaaga
     1801 aacaaactga accaagagca gaatgccaag ctgcaacaac agagggagtg tttgaataag
     1861 cgcaattcag aggtggccgt catggataaa cgcgtcagtg agctgaggga ccggctgtgg
     1921 aagaagaagg cagcactgca gcagaaagag aatctcccgg tttcacctga tggaaatctt
     1981 ccccaacaag cagtgtcagc cccaagtcgt gtggctgctg taggccctta catccagtca
     2041 tccactatgc cacggatgcc ctccagacct gagctgctcg tgaagccagc cctgcctgat
     2101 ggttccttgc tcatgcagtc agcagagggg ccaatgaaga tacagacact gcccaatatg
     2161 aggtctgggg ctgcttcaca aagtaaaggc tctaaagctc acccagctag ccccgattgg
     2221 aacccttcca atgccgacct cttacccagc caaggctctt ctgtacccca gagtgctgga
     2281 actgctctgg accaagttga cgatggggag attgctgtga gggagaaaga gaagaaagtg
     2341 cgtccctttt ccatgtttga cacagtggac cagtgtgctg ccccaccctc ctttggtacc
     2401 ctgaggaaaa accagagcag cgaggacatc ttgcgggatg ctcaggctgt aaataaaaat
     2461 gtagctaaag taccacctcc cgttcctaca aagccaaaac agattcattt gccttacttt
     2521 ggacaaacgg ctcagtcgcc ttctgacatg aagccagatg gaaatgctca gcaattgcca
     2581 atagctgcta catccgtggg ggctaagctc aagccagcag ggccacaggc aaggatgctg
     2641 ctgtctcctg gtgccccttc aggtggtcag gaccaagtcc tgtctccagc atctaagcaa
     2701 gaaagtcctc ctgctgctgc agtccggccc ttcacgcccc agccgtccaa ggacaccttc
     2761 cctccagcct ttcgaaaacc ccagactgtg gcagcaagct ccatctattc catgtacacc
     2821 cagcaacagg caccaggaaa aaacttccag caggctgtgc agagtgcctt gaccaagacg
     2881 caacccagag gcccccactt ttcaagtgtg tatggcaagc ctgtgatagc tgctgcccag
     2941 aatccacaac agcacccaga gaacatttac tcctgtagcc aggggaagcc tggcagtccg
     3001 gagcccgaga cagagaccgt ttcttcagtt catgagagcc atgaaaacga aaggattcct
     3061 cgaccactca gcccaacaaa attactgcct ttcttgtcta acccttatcg aaaccagagc
     3121 gatgctgact tagaagccct gaggaaaaaa ttgtcgaatg caccacggcc actaaagaaa
     3181 cgtagctcca ttacagaacc agaaggccct aatgggccaa atattcagaa acttttgtat
     3241 cagaggacca ccatagctgc catggagacc atctcagtcc cctcacaccc atctaagtca
     3301 ccaggctcag tgactgtcaa tccagagagc tccgttgaaa tcccaaatcc atatttacat
     3361 gtggagccag agaaggaggt gggctcttta gtccctgaac cactgtcccc agaggatatg
     3421 gggagtgcca gtacggagaa cagtgacgtg ccagctcctt ctgcaggcct tgagtatgtg
     3481 tctgagggag tcacagacag cagtaccaat ctgcagaata acgtagaaga aacaaaccca
     3541 gaggctcccc acttacttga agtgtaccta gaagaatacc ccccataccc accccctcca
     3601 tacccatctg gggagccaga ggtgtccgaa gaagactccg cacgtatgcg cccacctgaa
     3661 atcaccgggc aggtgtctct gcctcctggc aaaaggacaa acttgcggaa aactggctcc
     3721 gagcggattg ctcacgggat gagggtgaaa ttcaaccccc ttgctttgct gctagattca
     3781 tctctggagg gagagtttga tcttgtacag agaattatct atgaggttga tgacccaagc
     3841 ctaccaaatg atgagggcat cacagctctt cataatgctg tgtgtgcagg ccacacagaa
     3901 attgtgaagt tcctggtaca gttcggtgtg aatgtaaatg cagcagatag tgatggatgg
     3961 acgccattgc attgtgctgc ctcctgcaac aatgtccaag tgtgtaagtt cctggtggag
     4021 tctggagcag ctgtgtttgc tatgacctac agtgacatgc agactgctgc agacaagtgc
     4081 gaggaaatgg aggaaggcta cacgcagtgc tcccagtttc tctatggcgt tcaggagaag
     4141 atgggcataa tgaacaaagg cgtcatttat gcactgtggg attacgagcc gcagcatgac
     4201 gatgagctgc tcatgaaaga aggagattgc atgaccgtca tccgcagaga agatgaagag
     4261 gagatcgagt ggtggtgggc gcgccttaac gataaggaag gatatgttcc acgcaacttg
     4321 ctaggactat acccaagaat taaaccaaga caaaggagct tggccctcga ggactacaaa
     4381 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     4441 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4501 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4561 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4621 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4681 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4741 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4801 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4861 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4921 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4981 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5041 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5101 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5161 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5221 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5281 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5341 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5401 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5461 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5521 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5581 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5641 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5701 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5761 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5821 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5881 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5941 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6001 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6061 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6121 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6181 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6241 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6301 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6361 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6421 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6481 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6541 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6661 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7561 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     7681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
