LOCUS       V020638                 9416 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020638
VERSION     V020638
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             683..4072
                     /label="LATS1(NM_004690)"
                     /note="LATS1(NM_004690)"
                     /gene="LATS1"
     CDS             4079..4168
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4192..4780
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4846..4894
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5096..5524
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5538..5867
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5934..6728
                     /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    6904..7025
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7074..7090)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7098..7114
                     /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(7122..7152)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7167..7188
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7476..8064)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8235..9095)
                     /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(9096..9200)
                     /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 ggatccgcca ccatgaagag gagtgaaaag ccagaaggat atagacaaat
      721 gaggcctaag acctttcctg ccagtaacta tactgtcagt agccggcaaa tgttacaaga
      781 aattcgggaa tcccttagga atttatctaa accatctgat gctgctaagg ctgagcataa
      841 catgagtaaa atgtcaaccg aagatcctcg acaagtcaga aatccaccca aatttgggac
      901 gcatcataaa gccttgcagg aaattcgaaa ctctctgctt ccatttgcaa atgaaacaaa
      961 ttcttctcgg agtacttcag aagttaatcc acaaatgctt caagacttgc aagctgctgg
     1021 atttgatgag gatatggtta tacaagctct tcagaaaact aacaacagaa gtatagaagc
     1081 agcaattgaa ttcattagta aaatgagtta ccaagatcct cgacgagagc agatggctgc
     1141 agcagctgcc agacctatta atgccagcat gaaaccaggg aatgtgcagc aatcagttaa
     1201 ccgcaaacag agctggaaag gttctaaaga atccttagtt cctcagaggc atggcccgcc
     1261 actaggagaa agtgtggcct atcattctga gagtcccaac tcacagacag atgtaggaag
     1321 acctttgtct ggatctggta tatcagcatt tgttcaagct caccctagca acggacagag
     1381 agtgaacccc ccaccaccac ctcaagtaag gagtgttact cctccaccac ctccaagagg
     1441 ccagactccc cctccaagag gtacaactcc acctccccct tcatgggaac caaactctca
     1501 aacaaagcgc tattctggaa acatggaata cgtaatctcc cgaatctctc ctgtcccacc
     1561 tggggcatgg caagagggct atcctccacc acctctcaac acttccccca tgaatcctcc
     1621 taatcaagga cagagaggca ttagttctgt tcctgttggc agacaaccaa tcatcatgca
     1681 gagttctagc aaatttaact ttccatcagg gagacctgga atgcagaatg gtactggaca
     1741 aactgatttc atgatacacc aaaatgttgt ccctgctggc actgtgaatc ggcagccacc
     1801 acctccatat cctctgacag cagctaatgg acaaagccct tctgctttac aaacaggggg
     1861 atctgctgct ccttcgtcat atacaaatgg aagtattcct cagtctatga tggtgccaaa
     1921 cagaaatagt cataacatgg aactatataa cattagtgta cctggactgc aaacaaattg
     1981 gcctcagtca tcttctgctc cagcccagtc atccccgagc agtgggcatg aaatccctac
     2041 atggcaacct aacataccag tgaggtcaaa ttcttttaat aacccattag gaaatagagc
     2101 aagtcactct gctaattctc agccttctgc tacaacagtc actgcaatta caccagctcc
     2161 tattcaacag cctgtgaaaa gtatgcgtgt attaaaacca gagctacaga ctgctttagc
     2221 acctacacac ccttcttgga taccacagcc aattcaaact gttcaaccca gtccttttcc
     2281 tgagggaacc gcttcaaatg tgactgtgat gccacctgtt gctgaagctc caaactatca
     2341 aggaccacca ccaccctacc caaaacatct gctgcaccaa aacccatctg ttcctccata
     2401 cgagtcaatc agtaagccta gcaaagagga tcagccaagc ttgcccaagg aagatgagag
     2461 tgaaaagagt tatgaaaatg ttgatagtgg ggataaagaa aagaaacaga ttacaacttc
     2521 acctattact gttaggaaaa acaagaaaga tgaagagcga agggaatctc gtattcaaag
     2581 ttattctcct caagcattta aattctttat ggagcaacat gtagaaaatg tactcaaatc
     2641 tcatcagcag cgtctacatc gtaaaaaaca attagagaat gaaatgatgc gggttggatt
     2701 atctcaagat gcccaggatc aaatgagaaa gatgctttgc caaaaagaat ctaattacat
     2761 ccgtcttaaa agggctaaaa tggacaagtc tatgtttgtg aagataaaga cactaggaat
     2821 aggagcattt ggtgaagtct gtctagcaag aaaagtagat actaaggctt tgtatgcaac
     2881 aaaaactctt cgaaagaaag atgttcttct tcgaaatcaa gtcgctcatg ttaaggctga
     2941 gagagatatc ctggctgaag ctgacaatga atgggtagtt cgtctatatt attcattcca
     3001 agataaggac aatttatact ttgtaatgga ctacattcct gggggtgata tgatgagcct
     3061 attaattaga atgggcatct ttccagaaag tctggcacga ttctacatag cagaacttac
     3121 ctgtgcagtt gaaagtgttc ataaaatggg ttttattcat agagatatta aacctgataa
     3181 tattttgatt gatcgtgatg gtcatattaa attgactgac tttggcctct gcactggctt
     3241 cagatggaca cacgattcta agtactatca gagtggtgac catccacggc aagatagcat
     3301 ggatttcagt aatgaatggg gggatccctc aagctgtcga tgtggagaca gactgaagcc
     3361 attagagcgg agagctgcac gccagcacca gcgatgtcta gcacattctt tggttgggac
     3421 tcccaattat attgcacctg aagtgttgct acgaacagga tacacacagt tgtgtgattg
     3481 gtggagtgtt ggtgttattc tttttgaaat gttggtggga caacctcctt tcttggcaca
     3541 aacaccatta gaaacacaaa tgaaggttat caactggcaa acatctcttc acattccacc
     3601 acaagctaaa ctcagtcctg aagcttctga tcttattatt aaactttccc gaggacccga
     3661 agatcgctta ggcaagaatg gtgctgatga aataaaagct catccatttt ttaaaacaat
     3721 tgacttctcc agtgacctga gacagcagtc tgcttcatac attcctaaaa tcacacaccc
     3781 aacagataca tcaaattttg atcctgttga tcctgataaa ttatggagtg atgataacga
     3841 ggaagaaaat gtaaatgaca ctctcaatgg atggtataaa aatggaaagc atcctgaaca
     3901 tgcattctat gaatttacct tccgaaggtt ttttgatgac aatggctacc catataatta
     3961 tccgaagcct attgaatatg aatacattaa ttcacaaggc tcagagcagc agtcggatga
     4021 agatgatcaa aacacaggct cagagattaa aaatcgcgat ctagtatatg ttctcgagga
     4081 acaaaaacta atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga
     4141 gcaaaagtta atttccgagg aggacttata gtgagggccc gatatctcga caatcaacct
     4201 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4261 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4321 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4381 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4441 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4501 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4561 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     4621 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4681 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4741 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     4801 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4861 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4921 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4981 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5041 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     5101 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5161 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5221 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     5281 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5341 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5401 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5461 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5521 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5881 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5941 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6001 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6061 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6121 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6181 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6241 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6301 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6361 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6421 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6481 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6541 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6601 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6661 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6721 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     6781 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6841 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6901 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6961 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7021 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7081 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7141 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7201 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7261 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     7681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8341 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9121 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9181 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     9241 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     9301 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     9361 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
