LOCUS       V039783                 8860 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039783
VERSION     V039783
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..4380
                     /label="LATS1(NM_004690)"
                     /note="LATS1(NM_004690)"
                     /gene="LATS1"
     polyA_signal    4460..4684
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4730..5158
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5172..5501
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5568..6362
                     /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    6536..6657
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6706..6722)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6730..6746
                     /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(6754..6784)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6799..6820
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7108..7693)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7864..8724)
                     /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(8725..8829)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgaagagga gtgaaaagcc agaaggatat
     1021 agacaaatga ggcctaagac ctttcctgcc agtaactata ctgtcagtag ccggcaaatg
     1081 ttacaagaaa ttcgggaatc ccttaggaat ttatctaaac catctgatgc tgctaaggct
     1141 gagcataaca tgagtaaaat gtcaaccgaa gatcctcgac aagtcagaaa tccacccaaa
     1201 tttgggacgc atcataaagc cttgcaggaa attcgaaact ctctgcttcc atttgcaaat
     1261 gaaacaaatt cttctcggag tacttcagaa gttaatccac aaatgcttca agacttgcaa
     1321 gctgctggat ttgatgagga tatggttata caagctcttc agaaaactaa caacagaagt
     1381 atagaagcag caattgaatt cattagtaaa atgagttacc aagatcctcg acgagagcag
     1441 atggctgcag cagctgccag acctattaat gccagcatga aaccagggaa tgtgcagcaa
     1501 tcagttaacc gcaaacagag ctggaaaggt tctaaagaat ccttagttcc tcagaggcat
     1561 ggcccgccac taggagaaag tgtggcctat cattctgaga gtcccaactc acagacagat
     1621 gtaggaagac ctttgtctgg atctggtata tcagcatttg ttcaagctca ccctagcaac
     1681 ggacagagag tgaacccccc accaccacct caagtaagga gtgttactcc tccaccacct
     1741 ccaagaggcc agactccccc tccaagaggt acaactccac ctcccccttc atgggaacca
     1801 aactctcaaa caaagcgcta ttctggaaac atggaatacg taatctcccg aatctctcct
     1861 gtcccacctg gggcatggca agagggctat cctccaccac ctctcaacac ttcccccatg
     1921 aatcctccta atcaaggaca gagaggcatt agttctgttc ctgttggcag acaaccaatc
     1981 atcatgcaga gttctagcaa atttaacttt ccatcaggga gacctggaat gcagaatggt
     2041 actggacaaa ctgatttcat gatacaccaa aatgttgtcc ctgctggcac tgtgaatcgg
     2101 cagccaccac ctccatatcc tctgacagca gctaatggac aaagcccttc tgctttacaa
     2161 acagggggat ctgctgctcc ttcgtcatat acaaatggaa gtattcctca gtctatgatg
     2221 gtgccaaaca gaaatagtca taacatggaa ctatataaca ttagtgtacc tggactgcaa
     2281 acaaattggc ctcagtcatc ttctgctcca gcccagtcat ccccgagcag tgggcatgaa
     2341 atccctacat ggcaacctaa cataccagtg aggtcaaatt cttttaataa cccattagga
     2401 aatagagcaa gtcactctgc taattctcag ccttctgcta caacagtcac tgcaattaca
     2461 ccagctccta ttcaacagcc tgtgaaaagt atgcgtgtat taaaaccaga gctacagact
     2521 gctttagcac ctacacaccc ttcttggata ccacagccaa ttcaaactgt tcaacccagt
     2581 ccttttcctg agggaaccgc ttcaaatgtg actgtgatgc cacctgttgc tgaagctcca
     2641 aactatcaag gaccaccacc accctaccca aaacatctgc tgcaccaaaa cccatctgtt
     2701 cctccatacg agtcaatcag taagcctagc aaagaggatc agccaagctt gcccaaggaa
     2761 gatgagagtg aaaagagtta tgaaaatgtt gatagtgggg ataaagaaaa gaaacagatt
     2821 acaacttcac ctattactgt taggaaaaac aagaaagatg aagagcgaag ggaatctcgt
     2881 attcaaagtt attctcctca agcatttaaa ttctttatgg agcaacatgt agaaaatgta
     2941 ctcaaatctc atcagcagcg tctacatcgt aaaaaacaat tagagaatga aatgatgcgg
     3001 gttggattat ctcaagatgc ccaggatcaa atgagaaaga tgctttgcca aaaagaatct
     3061 aattacatcc gtcttaaaag ggctaaaatg gacaagtcta tgtttgtgaa gataaagaca
     3121 ctaggaatag gagcatttgg tgaagtctgt ctagcaagaa aagtagatac taaggctttg
     3181 tatgcaacaa aaactcttcg aaagaaagat gttcttcttc gaaatcaagt cgctcatgtt
     3241 aaggctgaga gagatatcct ggctgaagct gacaatgaat gggtagttcg tctatattat
     3301 tcattccaag ataaggacaa tttatacttt gtaatggact acattcctgg gggtgatatg
     3361 atgagcctat taattagaat gggcatcttt ccagaaagtc tggcacgatt ctacatagca
     3421 gaacttacct gtgcagttga aagtgttcat aaaatgggtt ttattcatag agatattaaa
     3481 cctgataata ttttgattga tcgtgatggt catattaaat tgactgactt tggcctctgc
     3541 actggcttca gatggacaca cgattctaag tactatcaga gtggtgacca tccacggcaa
     3601 gatagcatgg atttcagtaa tgaatggggg gatccctcaa gctgtcgatg tggagacaga
     3661 ctgaagccat tagagcggag agctgcacgc cagcaccagc gatgtctagc acattctttg
     3721 gttgggactc ccaattatat tgcacctgaa gtgttgctac gaacaggata cacacagttg
     3781 tgtgattggt ggagtgttgg tgttattctt tttgaaatgt tggtgggaca acctcctttc
     3841 ttggcacaaa caccattaga aacacaaatg aaggttatca actggcaaac atctcttcac
     3901 attccaccac aagctaaact cagtcctgaa gcttctgatc ttattattaa actttgccga
     3961 ggacccgaag atcgcttagg caagaatggt gctgatgaaa taaaagctca tccatttttt
     4021 aaaacaattg acttctccag tgacctgaga cagcagtctg cttcatacat tcctaaaatc
     4081 acacacccaa cagatacatc aaattttgat cctgttgatc ctgataaatt atggagtgat
     4141 gataacgagg aagaaaatgt aaatgacact ctcaatggat ggtataaaaa tggaaagcat
     4201 cctgaacatg cattctatga atttaccttc cgaaggtttt ttgatgacaa tggctaccca
     4261 tataattatc cgaagcctat tgaatatgaa tacattaatt cacaaggctc agagcagcag
     4321 tcggatgaag atgatcaaaa cacaggctca gagattaaaa atcgcgatct agtatatgtt
     4381 taagaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     4441 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4501 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4561 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4621 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4681 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4741 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4801 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4861 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4921 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4981 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5041 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5101 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     5161 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5221 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5281 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5341 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5401 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5461 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5521 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5581 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5641 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5701 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5761 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5821 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5881 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5941 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6001 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6061 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     6121 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6181 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6241 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6301 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6361 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6421 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6481 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6541 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6601 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6661 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6721 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6781 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6841 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6901 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6961 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7621 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7681 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7741 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7801 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7861 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7921 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7981 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8041 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8101 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8161 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8221 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8281 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8341 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8401 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8461 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8521 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8581 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8641 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8701 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8761 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8821 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
