LOCUS       V034760                 8788 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034760
VERSION     V034760
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..3444
                     /label="Cbl(NM_007619)"
                     /note="Cbl(NM_007619)"
                     /gene="Cbl"
     CDS             3451..3540
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3564..4152
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4218..4266
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4468..4896
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4910..5239
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5306..6100
                     /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    6276..6397
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6446..6462)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6470..6486
                     /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(6494..6524)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6539..6560
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6848..7436)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7607..8467)
                     /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(8468..8572)
                     /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 ccaccatggc cggcaacgtg
      721 aagaagagct cgggcgccgg cggcggcggc tctgggggct cgggagcggg cggcctgatc
      781 gggctcatga aggacgcctt ccagccgcac caccaccacc accacctcag cccgcaccct
      841 ccctgcacgg tggacaagaa gatggtggag aagtgctgga agctcatgga caaggtggtg
      901 cggttgtgtc aaaacccaaa gctggcgctc aagaacagcc cgccttatat cttagacctg
      961 ctgcctgaca cctaccagca cctccgcact gtcttgtcaa gatatgaggg gaagatggag
     1021 acgcttggag aaaatgagta tttcagggtg ttcatggaaa atttgatgaa gaaaactaag
     1081 cagactatca gcctcttcaa ggagggaaaa gaaaggatgt atgaggagaa ttcccagcct
     1141 aggcgaaacc tgaccaaatt atccctgatc ttcagccaca tgctggcaga actgaaaggc
     1201 atctttccga gcggactctt ccaaggagac actttccgga ttactaaagc tgatgctgcc
     1261 gaattttgga gaaaagcttt tggtgaaaag acgatagtcc cgtggaagag ctttcgacag
     1321 gccctgcatg aagtgcatcc catcagttct gggctggagg ccatggctct gaagtccact
     1381 attgatctga cctgcaatga ttatatttct gtctttgaat ttgatatttt tacacggctg
     1441 tttcagccct ggtcctcttt gctcagaaat tggaacagcc ttgctgtaac tcaccctggt
     1501 tacatggctt tcctgacata cgatgaagtg aaagcgcgcc tgcagaagtt catccacaaa
     1561 cctggcagtt acatctttcg gctgagctgt actcgtttgg gtcagtgggc tattgggtat
     1621 gttactgccg atgggaacat tctgcagaca atcccacaca ataaaccgct cttccaagca
     1681 ctgattgatg gcttcaggga aggcttctat ttgtttcctg atggacgaaa tcaaaatcct
     1741 gacctgacag gtttatgtga accaactcct caagatcata tcaaagtaac ccaggaacaa
     1801 tatgaattat actgtgaaat gggctccaca tttcaactgt gtaagatatg tgctgagaat
     1861 gataaggatg tgaagattga gccctgtgga cacctcatgt gcacatcctg cctcacgtcg
     1921 tggcaggaat cagaaggtca gggctgtcct ttttgccgat gtgaaatcaa aggtactgag
     1981 cccatcgtgg tggatccgtt tgaccccaga ggcagtggca gcctattaag gcaaggagca
     2041 gaaggtgctc cttccccaaa ttacgacgat gatgatgatg aacgagctga tgattctctc
     2101 ttcatgatga aggagttggc aggtgccaag gtggaaaggc cttcctctcc attctccatg
     2161 gccccacaag cttcccttcc tccagtgcca ccaagacttg accttctaca gcagcgagca
     2221 cctgttcctg ccagcacttc agttctgggg actgcttcca aggctgcttc tggctccctt
     2281 cataaagaca aaccattgcc aatacctccc acacttcgag atcttccacc accaccccct
     2341 ccagaccggc cttactctgt tggagcagaa acaaggcctc agagacgccc tctgccttgt
     2401 acaccaggcg attgtccatc tagagacaaa ctgccccctg tcccttctag ccgcccaggg
     2461 gactcgtggt tgtctcggcc aatccctaaa gtaccagtag ctactccaaa ccctggtgat
     2521 ccttggaatg ggagagaatt gaccaatcgg cactcgcttc cattctcatt gccctcacaa
     2581 atggaaccca gagcagatgt ccctaggctt ggaagcacat ttagtctgga tacctctatg
     2641 actatgaata gcagcccagt agcaggtcca gagagtgagc acccaaagat caagccttcc
     2701 tcgtctgcca acgccattta ctctctggct gccaggcctc ttcctatgcc aaaactgcca
     2761 cctggggagc aaggggaaag tgaagaggac acagaatata tgactcccac atctaggcct
     2821 gtaggggttc agaagccaga gcccaaacgg ccgttagagg caacccagag ttcacgagca
     2881 tgtgactgtg accagcagat cgacagctgt acctatgaag cgatgtataa catccagtcc
     2941 caagcactgt ctgtagcaga aaacagcgcc tctggggaag ggaatctggc cacagctcac
     3001 acgagtactg gccctgagga atccgaaaac gaggatgatg gctatgatgt gcctaagcca
     3061 cccgtgccag ctgtactggc ccgccggacc ctgtctgaca tctccaatgc cagctcctcc
     3121 tttggctggt tgtctttgga tggtgaccct acaaacttca atgagggttc ccaagttcct
     3181 gagcggcccc ccaaaccatt ccctcggaga atcaactcag aacgaaaagc cagtagctat
     3241 caacaaggcg gaggtgccac tgctaaccct gtggccacag caccctcacc gcagctctca
     3301 agtgagattg aacgcctcat gagtcagggc tattcctacc aggacattca gaaagctttg
     3361 gtcattgccc acaacaacat tgagatggct aaaaacatcc tccgggaatt tgtttctatt
     3421 tcttctcctg ctcacgtagc caccctcgag gaacaaaaac taatatcaga ggaagatttg
     3481 gaacagaaac taatcagtga agaagattta gagcaaaagt taatttccga ggaggactta
     3541 tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     3601 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3661 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3721 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3781 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3841 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3901 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     3961 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4021 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4081 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4141 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     4201 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     4261 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4321 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     4381 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     4441 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4501 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4561 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4621 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4681 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4741 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4801 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     4861 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4921 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4981 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5041 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5101 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5161 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5221 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5281 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5341 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5401 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5461 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5521 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5581 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5641 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5701 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5761 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5821 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5881 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5941 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6001 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6061 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     6121 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     6181 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6241 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6301 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6421 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6601 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7021 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     7081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7621 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7681 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7741 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7801 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7861 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7921 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7981 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8041 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8101 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8161 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8221 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8281 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8341 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8401 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8461 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8521 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8581 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     8641 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     8701 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     8761 cgacaattgc atgaagaatc tgcttagg
//
