LOCUS       V024158                 7079 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024158
VERSION     V024158
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             941..2569
                     /label="YES1(NM_005433)"
                     /note="YES1(NM_005433)"
                     /gene="YES1"
     CDS             2576..2641
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2679..2903
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2949..3377
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3391..3720
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3787..4581
                     /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    4755..4876
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4925..4941)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4949..4965
                     /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(4973..5003)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5018..5039
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5327..5912)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6083..6943)
                     /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(6944..7048)
                     /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 atccgccacc atgggctgca ttaaaagtaa
      961 agaaaacaaa agtccagcca ttaaatacag acctgaaaat actccagagc ctgtcagtac
     1021 aagtgtgagc cattatggag cagaacccac tacagtgtca ccatgtccgt catcttcagc
     1081 aaagggaaca gcagttaatt tcagcagtct ttccatgaca ccatttggag gatcctcagg
     1141 ggtaacgcct tttggaggtg catcttcctc attttcagtg gtgccaagtt catatcctgc
     1201 tggtttaaca ggtggtgtta ctatatttgt ggccttatat gattatgaag ctagaactac
     1261 agaagacctt tcatttaaga agggtgaaag atttcaaata attaacaata cggaaggaga
     1321 ttggtgggaa gcaagatcaa tcgctacagg aaagaatggt tatatcccga gcaattatgt
     1381 agcgcctgca gattccattc aggcagaaga atggtatttt ggcaaaatgg ggagaaaaga
     1441 tgctgaaaga ttacttttga atcctggaaa tcaacgaggt attttcttag taagagagag
     1501 tgaaacaact aaaggtgctt attccctttc tattcgtgat tgggatgaga taaggggtga
     1561 caatgtgaaa cactacaaaa ttaggaaact tgacaatggt ggatactata tcacaaccag
     1621 agcacaattt gatactctgc agaaattggt gaaacactac acagaacatg ctgatggttt
     1681 atgccacaag ttgacaactg tgtgtccaac tgtgaaacct cagactcaag gtctagcaaa
     1741 agatgcttgg gaaatccctc gagaatcttt gcgactagag gttaaactag gacaaggatg
     1801 tttcggcgaa gtgtggatgg gaacatggaa tggaaccacg aaagtagcaa tcaaaacact
     1861 aaaaccaggt acaatgatgc cagaagcttt ccttcaagaa gctcagataa tgaaaaaatt
     1921 aagacatgat aaacttgttc cactatatgc tgttgtttct gaagaaccaa tttacattgt
     1981 cactgaattt atgtcaaaag gaagcttatt agatttcctt aaggaaggag atggaaagta
     2041 tttgaagctt ccacagctgg ttgatatggc tgctcagatt gctgatggta tggcatatat
     2101 tgaaagaatg aactatattc accgagatct tcgggctgct aatattcttg taggagaaaa
     2161 tcttgtgtgc aaaatagcag actttggttt agcaaggtta attgaagaca atgaatacac
     2221 agcaagacaa ggtgcaaaat ttccaatcaa atggacagct cctgaagctg cactgtatgg
     2281 tcggtttaca ataaagtctg atgtctggtc atttggaatt ctgcaaacag aactagtaac
     2341 aaagggccga gtgccatatc caggtatggt gaaccgtgaa gtattagaac aagtggagcg
     2401 aggatacagg atgccgtgcc ctcagggctg tccagaatcc ctccatgaat tgatgaatct
     2461 gtgttggaag aaggaccctg atgaaagacc aacatttgaa tatattcagt ccttcttgga
     2521 agactacttc actgctacag agccacagta ccagccagga gaaaatttac tcgaggacta
     2581 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     2641 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2701 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2761 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2821 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2881 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2941 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3001 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3061 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3121 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3181 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3241 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3301 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3361 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3421 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3541 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3601 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3661 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3721 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3781 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3841 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3901 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3961 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4021 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4081 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4141 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4201 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4261 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4321 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4381 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4441 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4501 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4561 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4621 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4681 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4741 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4801 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4861 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4921 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4981 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5041 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5101 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5161 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5221 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5281 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5341 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5401 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5461 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5521 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5581 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5641 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5701 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5761 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5821 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     5881 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5941 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
