LOCUS       V029009                 7728 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029009
VERSION     V029009
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..2408
                     /label="TRAF3IP3(NM_025228)"
                     /note="TRAF3IP3(NM_025228)"
                     /gene="TRAF3IP3"
     CDS             2415..2480
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2504..3092
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3158..3206
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3408..3836
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3850..4179
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4246..5040
                     /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    5216..5337
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5386..5402)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5410..5426
                     /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(5434..5464)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5479..5500
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5788..6376)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6547..7407)
                     /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(7408..7512)
                     /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 ccaccatgat cagcccagac cccaggccct
      781 cccctggctt ggcccggtgg gctgagagct atgaggccaa gtgtgagcgc aggcaagaga
      841 tccgtgaaag ccgccgctgc cgtcccaatg tgaccacttg ccgccaggtg gggaagacgc
      901 tgaggatcca acagagagag cagctccaga gagctcgact gcagcagttc ttcaggagga
      961 ggaacctgga gctagaggag aagggcaaag cgcagcatcc ccaggccagg gagcaagggc
     1021 cctccaggcg gccaggacag gtgactgtcc tcaaggaacc cttgtcttgt gccagaagga
     1081 tttcttctcc cagagagcag gtgacaggca ccagctctga agtctttcca gcccagcatc
     1141 ctcctccctc aggcatctgc agggatctgt ctgaccacct ctcctcacag gctgggggcc
     1201 ttcctccaca ggacactccc atcaagaagc cacccaaaca ccaccgtggt actcagacaa
     1261 aggcagaagg accaacaatt aagaacgatg ccagtcagca aaccaattac ggagttgcag
     1321 ttctggataa ggaaatcatc cagctttctg attacctcaa agaggcccta caaagggagc
     1381 tggtcctaaa acagaaaatg gtgattctcc aagacctact gtccactctg attcaggcct
     1441 ctgacagctc ttggaaggga cagcttaatg aagacaaact gaaggggaaa ctgagatcct
     1501 tagaaaacca gctatacacc tgtacccaga aatactcccc ttggggaatg aaaaaagtac
     1561 tactggagat ggaagaccag aaaaacagct atgagcagaa ggccaaggag tcactgcaga
     1621 aagtgctgga ggagaaaatg aatgcagagc agcaactaca gagcacacag cgatccctgg
     1681 ccctggcaga gcagaagtgt gaagagtgga ggagccagta tgaggctctg aaggaggact
     1741 ggaggaccct tgggacccag cacagggagc tggagagcca actccacgtg cttcagtcca
     1801 aactgcaggg agcagatagc agggacttac agatgaacca ggccctgcga tttttggaaa
     1861 atgagcacca gcaactgcag gccaagattg aatgcctgca aggggacaga gacctgtgca
     1921 gcttggatac ccaggaccta caagatcaac taaaaaggtc agaggcagag aaactcaccc
     1981 tggtgaccag agtacagcag ttgcagggtt tgcttcaaaa tcaatcctta cagcttcaag
     2041 aacaggagaa actcttaaca aagaaagatc aggctttgcc cgtgtggagt ccaaagtcct
     2101 tccctaacga agtggagcct gagggtacag ggaaggagaa agactgggat ctcagagacc
     2161 agctgcaaaa gaagactttg cagctccagg ccaaggaaaa ggagtgcaga gaactgcatt
     2221 cagaattaga caacctcagt gacgagtatc tctcctgcct gcgtaagctg cagcactgtc
     2281 gagaagagct gaaccagagc cagcagctgc ctcccagaag gcaatgtggg cgatggctcc
     2341 cagtgctgat ggtggtgatt gctgcagcac tggcagtgtt cctggccaat aaagacaacc
     2401 tgatgatcgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2461 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2521 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2581 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2641 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2701 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2761 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2821 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2881 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2941 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3001 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3061 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3121 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3181 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3241 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3301 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3361 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3421 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3481 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3541 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3601 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3661 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3721 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3781 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3841 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4021 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4081 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4141 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4201 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4261 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4321 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4381 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4441 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4501 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4561 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4621 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4681 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4741 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4801 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4861 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4921 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4981 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5041 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5101 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5161 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5221 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5281 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5341 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7561 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7621 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7681 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
