LOCUS       V035571                 8861 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035571
VERSION     V035571
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..4297
                     /label="LATS1(NM_004690)"
                     /note="LATS1(NM_004690)"
                     /gene="LATS1"
     CDS             4304..4393
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    4428..4652
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4698..5126
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5140..5469
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5536..6330
                     /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    6504..6625
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6674..6690)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6698..6714
                     /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(6722..6752)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6767..6788
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7076..7661)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7832..8692)
                     /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(8693..8797)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg aagaggagtg aaaagccaga aggatataga caaatgaggc ctaagacctt
      961 tcctgccagt aactatactg tcagtagccg gcaaatgtta caagaaattc gggaatccct
     1021 taggaattta tctaaaccat ctgatgctgc taaggctgag cataacatga gtaaaatgtc
     1081 aaccgaagat cctcgacaag tcagaaatcc acccaaattt gggacgcatc ataaagcctt
     1141 gcaggaaatt cgaaactctc tgcttccatt tgcaaatgaa acaaattctt ctcggagtac
     1201 ttcagaagtt aatccacaaa tgcttcaaga cttgcaagct gctggatttg atgaggatat
     1261 ggttatacaa gctcttcaga aaactaacaa cagaagtata gaagcagcaa ttgaattcat
     1321 tagtaaaatg agttaccaag atcctcgacg agagcagatg gctgcagcag ctgccagacc
     1381 tattaatgcc agcatgaaac cagggaatgt gcagcaatca gttaaccgca aacagagctg
     1441 gaaaggttct aaagaatcct tagttcctca gaggcatggc ccgccactag gagaaagtgt
     1501 ggcctatcat tctgagagtc ccaactcaca gacagatgta ggaagacctt tgtctggatc
     1561 tggtatatca gcatttgttc aagctcaccc tagcaacgga cagagagtga accccccacc
     1621 accacctcaa gtaaggagtg ttactcctcc accacctcca agaggccaga ctccccctcc
     1681 aagaggtaca actccacctc ccccttcatg ggaaccaaac tctcaaacaa agcgctattc
     1741 tggaaacatg gaatacgtaa tctcccgaat ctctcctgtc ccacctgggg catggcaaga
     1801 gggctatcct ccaccacctc tcaacacttc ccccatgaat cctcctaatc aaggacagag
     1861 aggcattagt tctgttcctg ttggcagaca accaatcatc atgcagagtt ctagcaaatt
     1921 taactttcca tcagggagac ctggaatgca gaatggtact ggacaaactg atttcatgat
     1981 acaccaaaat gttgtccctg ctggcactgt gaatcggcag ccaccacctc catatcctct
     2041 gacagcagct aatggacaaa gcccttctgc tttacaaaca gggggatctg ctgctccttc
     2101 gtcatataca aatggaagta ttcctcagtc tatgatggtg ccaaacagaa atagtcataa
     2161 catggaacta tataacatta gtgtacctgg actgcaaaca aattggcctc agtcatcttc
     2221 tgctccagcc cagtcatccc cgagcagtgg gcatgaaatc cctacatggc aacctaacat
     2281 accagtgagg tcaaattctt ttaataaccc attaggaaat agagcaagtc actctgctaa
     2341 ttctcagcct tctgctacaa cagtcactgc aattacacca gctcctattc aacagcctgt
     2401 gaaaagtatg cgtgtattaa aaccagagct acagactgct ttagcaccta cacacccttc
     2461 ttggatacca cagccaattc aaactgttca acccagtcct tttcctgagg gaaccgcttc
     2521 aaatgtgact gtgatgccac ctgttgctga agctccaaac tatcaaggac caccaccacc
     2581 ctacccaaaa catctgctgc accaaaaccc atctgttcct ccatacgagt caatcagtaa
     2641 gcctagcaaa gaggatcagc caagcttgcc caaggaagat gagagtgaaa agagttatga
     2701 aaatgttgat agtggggata aagaaaagaa acagattaca acttcaccta ttactgttag
     2761 gaaaaacaag aaagatgaag agcgaaggga atctcgtatt caaagttatt ctcctcaagc
     2821 atttaaattc tttatggagc aacatgtaga aaatgtactc aaatctcatc agcagcgtct
     2881 acatcgtaaa aaacaattag agaatgaaat gatgcgggtt ggattatctc aagatgccca
     2941 ggatcaaatg agaaagatgc tttgccaaaa agaatctaat tacatccgtc ttaaaagggc
     3001 taaaatggac aagtctatgt ttgtgaagat aaagacacta ggaataggag catttggtga
     3061 agtctgtcta gcaagaaaag tagatactaa ggctttgtat gcaacaaaaa ctcttcgaaa
     3121 gaaagatgtt cttcttcgaa atcaagtcgc tcatgttaag gctgagagag atatcctggc
     3181 tgaagctgac aatgaatggg tagttcgtct atattattca ttccaagata aggacaattt
     3241 atactttgta atggactaca ttcctggggg tgatatgatg agcctattaa ttagaatggg
     3301 catctttcca gaaagtctgg cacgattcta catagcagaa cttacctgtg cagttgaaag
     3361 tgttcataaa atgggtttta ttcatagaga tattaaacct gataatattt tgattgatcg
     3421 tgatggtcat attaaattga ctgactttgg cctctgcact ggcttcagat ggacacacga
     3481 ttctaagtac tatcagagtg gtgaccatcc acggcaagat agcatggatt tcagtaatga
     3541 atggggggat ccctcaagct gtcgatgtgg agacagactg aagccattag agcggagagc
     3601 tgcacgccag caccagcgat gtctagcaca ttctttggtt gggactccca attatattgc
     3661 acctgaagtg ttgctacgaa caggatacac acagttgtgt gattggtgga gtgttggtgt
     3721 tattcttttt gaaatgttgg tgggacaacc tcctttcttg gcacaaacac cattagaaac
     3781 acaaatgaag gttatcaact ggcaaacatc tcttcacatt ccaccacaag ctaaactcag
     3841 tcctgaagct tctgatctta ttattaaact ttgccgagga cccgaagatc gcttaggcaa
     3901 gaatggtgct gatgaaataa aagctcatcc attttttaaa acaattgact tctccagtga
     3961 cctgagacag cagtctgctt catacattcc taaaatcaca cacccaacag atacatcaaa
     4021 ttttgatcct gttgatcctg ataaattatg gagtgatgat aacgaggaag aaaatgtaaa
     4081 tgacactctc aatggatggt ataaaaatgg aaagcatcct gaacatgcat tctatgaatt
     4141 taccttccga aggttttttg atgacaatgg ctacccatat aattatccga agcctattga
     4201 atatgaatac attaattcac aaggctcaga gcagcagtcg gatgaagatg atcaaaacac
     4261 aggctcagag attaaaaatc gcgatctagt atatgttctc gaggaacaaa aactaatatc
     4321 agaggaagat ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc
     4381 cgaggaggac ttatgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     4441 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4501 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4561 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4621 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4681 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4741 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4801 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4861 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4921 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4981 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5041 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5101 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5161 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5221 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5281 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5341 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5401 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5461 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5521 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5581 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5641 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5701 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5761 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5821 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5881 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5941 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6001 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6061 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6121 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6181 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6241 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6301 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6361 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6421 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6481 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6541 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6601 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6661 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6721 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6781 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6841 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6901 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6961 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7021 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7081 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7141 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7201 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7261 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7321 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7381 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7441 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7501 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7561 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     7621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8821 ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
