LOCUS       V031643                 7672 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031643
VERSION     V031643
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..2328
                     /label="Src(NM_009271)"
                     /note="Src(NM_009271)"
                     /gene="Src"
     CDS             2335..2424
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2448..3036
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3102..3150
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3352..3780
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3794..4123
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4190..4984
                     /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    5160..5281
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5330..5346)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5354..5370
                     /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(5378..5408)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5423..5444
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5732..6320)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6491..7351)
                     /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(7352..7456)
                     /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 ccaccatggg cagcaacaag
      721 agcaagccca aggacgccag ccagcggcgc cgcagcctgg agccctcgga aaacgtgcac
      781 ggggcagggg gcgccttccc ggcctcacag acaccgagca agcccgcctc cgccgacggc
      841 caccgcgggc ccagcgccgc cttcgtgccg cccgcggccg agcccaagct cttcggaggc
      901 ttcaactcct cggacaccgt cacctccccg cagagggcgg ggcctctggc aggtggggtg
      961 accacctttg tggccctcta tgactatgag tcacggacag agactgacct gtccttcaag
     1021 aaaggggagc ggctgcagat tgtcaataac acgaggaagg tggatgtcag agagggagac
     1081 tggtggctgg cacactcgct gagcacggga cagaccggtt acatccccag caactatgtg
     1141 gcgccctccg actccatcca ggctgaggag tggtactttg gcaagatcac tagacgggaa
     1201 tcagagcggc tgctgctcaa cgccgagaac ccgagaggga ccttcctcgt gagggagagt
     1261 gagaccacaa aaggtgccta ctgcctctct gtatccgact tcgacaatgc caagggccta
     1321 aatgtgaaac actacaagat ccgcaagctg gacagcggcg gtttctacat cacctcccgc
     1381 acccagttca acagcctgca gcagctcgtg gcttactact ccaaacatgc tgatggcctg
     1441 tgtcaccgcc tcactaccgt atgtcccaca tccaagcctc agacccaggg attggccaag
     1501 gatgcgtggg agatcccccg ggagtccctg cggctggagg tcaagctggg ccagggttgc
     1561 ttcggagagg tgtggatggg gacctggaac ggcaccacga gggttgccat caaaactctg
     1621 aagccaggca ccatgtcccc agaggccttc ctgcaggagg cccaagtcat gaagaaactg
     1681 aggcacgaga aactggtgca gctgtatgct gtggtgtcgg aagaacccat ttacattgtg
     1741 acagagtaca tgaacaaggg gagtctgctg gactttctca agggggaaac gggcaaatat
     1801 ttgcggctac cccagctggt ggacatgtct gctcagatcg cttcaggcat ggcctatgtg
     1861 gagcggatga actatgtgca ccgggacctt cgagccgcca atatcctagt aggggagaac
     1921 ctggtgtgca aagtggccga ctttgggttg gcccggctca tagaagacaa cgaatacaca
     1981 gcccggcaag gtgccaaatt ccccatcaag tggaccgccc ctgaagctgc tctgtacggc
     2041 aggttcacca tcaagtcgga tgtgtggtcc tttgggattc tgctgaccga gctcaccact
     2101 aagggaagag tgccctatcc tgggatggtg aaccgtgagg ttctggacca ggtggagcgg
     2161 ggctaccgga tgccttgtcc ccccgagtgc cccgagtccc tgcatgacct tatgtgccag
     2221 tgctggcgga aggagcccga ggagcggccc accttcgagt acctgcaggc cttcctggaa
     2281 gactacttta cgtccactga gccacagtac cagcccgggg agaacctact cgaggaacaa
     2341 aaactaatat cagaggaaga tttggaacag aaactaatca gtgaagaaga tttagagcaa
     2401 aagttaattt ccgaggagga cttatagtga gggcccgata tctcgacaat caacctctgg
     2461 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2521 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2581 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2641 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     2701 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     2761 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2821 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     2881 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     2941 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3001 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga
     3061 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3121 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     3181 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     3241 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     3301 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
     3361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3421 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3481 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     3541 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3661 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3721 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3781 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3841 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3901 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3961 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4021 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4081 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4141 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4201 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4261 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4321 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4381 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4441 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4501 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4561 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4621 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4681 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4741 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4801 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4861 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4921 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4981 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5041 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5101 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5281 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5461 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     5941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7501 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag
     7561 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7621 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
