LOCUS       V042753                 7639 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042753
VERSION     V042753
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..2295
                     /label="Traf6(NM_009424)"
                     /note="Traf6(NM_009424)"
                     /gene="Traf6"
     CDS             2302..2391
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2415..3003
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3069..3117
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3319..3747
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3761..4090
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4157..4951
                     /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    5127..5248
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5297..5313)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5321..5337
                     /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(5345..5375)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5390..5411
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5699..6287)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6458..7318)
                     /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(7319..7423)
                     /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 ccaccatgag tctcttaaac
      721 tgtgagaaca gctgcgggtc cagccagtcg tccagtgact gctgcgctgc catggccgcc
      781 tcctgcagcg ctgcagtgaa agatgacagc gtgagtggct ctgccagcac cgggaacctc
      841 tccagctcct tcatggagga gatccagggc tacgatgtgg agtttgaccc acctctggag
      901 agcaagtatg agtgtcccat ctgcttgatg gctttacggg aagcagtgca aacaccatgt
      961 ggccacaggt tctgcaaagc ctgcatcatc aaatccataa gggatgcagg gcacaagtgc
     1021 ccagttgaca atgaaatact gctggaaaat caactgtttc ccgacaattt tgcaaagcga
     1081 gagattcttt ccctgacggt aaagtgccca aataaaggct gtttgcaaaa gatggaactg
     1141 agacatctcg aggatcatca agtacattgt gaatttgctc tagtgaattg tccccagtgc
     1201 caacgtcctt tccagaagtg ccaggttaat acacacatta ttgaggattg tcccaggagg
     1261 caggtttctt gtgtaaactg tgctgtgtcc atggcatatg aagagaaaga gatccatgat
     1321 caaagctgtc ctctggcaaa tatcatctgt gaatactgtg gtacaatcct catcagagaa
     1381 cagatgccta atcattatga tctggactgc ccaacagctc caatcccttg cacattcagt
     1441 gtttttggct gtcatgaaaa gatgcagagg aatcacttgg cacgacactt gcaagagaat
     1501 acccagttgc acatgagact gttggcccag gctgttcata atgttaacct tgctttgcgt
     1561 ccgtgcgatg ccgcctctcc atcccgggga tgtcgtccag aggacccaaa ttatgaggaa
     1621 actatcaaac agttggagag tcgcctagta agacaggacc atcagatccg ggagctgact
     1681 gccaaaatgg aaactcagag tatgtacgtg ggcgagctca aacggaccat tcggaccctg
     1741 gaggacaagg ttgccgaaat ggaagcacag cagtgtaacg ggatctacat ttggaagatt
     1801 ggcaactttg ggatgcactt gaaatcccaa gaagaggaaa gacctgttgt catccatagc
     1861 cctggattct acacaggcag acctgggtac aagctgtgca tgcgcctgca tcttcagtta
     1921 ccgacagctc agcgctgtgc aaactatata tccctttttg tccacacaat gcaaggagaa
     1981 tatgacagcc acctcccctg gcccttccag ggtacaatac gccttacaat tctcgaccag
     2041 tctgaagcac ttataaggca aaaccacgaa gaggtcatgg acgccaaacc agaactgctt
     2101 gcctttcagc gacccacaat cccacggaac cccaaaggtt ttggctatgt aacatttatg
     2161 cacctggaag ccttaagaca gggaaccttc attaaggatg atacattact agtgcgctgt
     2221 gaagtctcta cccgctttga catgggtggc cttcggaagg agggtttcca gccacgaagt
     2281 actgatgcgg gggtgctcga ggaacaaaaa ctaatatcag aggaagattt ggaacagaaa
     2341 ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt atagtgaggg
     2401 cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2461 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2521 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2581 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2641 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2701 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2761 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     2821 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2881 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2941 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3001 cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3061 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     3121 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3181 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3241 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg
     3301 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3361 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3421 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     3481 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3541 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3601 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3661 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     3721 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3781 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3841 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3961 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4021 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4081 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4141 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4201 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4261 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4321 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4381 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4441 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4501 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4561 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4621 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4681 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4741 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4801 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4861 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4921 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     4981 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5041 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5101 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5161 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5221 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5281 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5341 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5401 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5461 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5521 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5581 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5641 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5701 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5761 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5821 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5881 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5941 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6001 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6061 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6121 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6181 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6301 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6361 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6421 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6481 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6541 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6601 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6661 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6721 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6781 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6841 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6901 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6961 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7021 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7081 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7141 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7201 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7261 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7321 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7381 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7441 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac
     7501 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     7561 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     7621 catgaagaat ctgcttagg
//
