LOCUS       V027469                 8049 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027469
VERSION     V027469
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..2775
                     /label="LATS1(NM_001270519)"
                     /note="LATS1(NM_001270519)"
                     /gene="LATS1"
     CDS             2782..2805
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    2825..3413
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3479..3527
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3729..4157
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4171..4500
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4567..5361
                     /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    5537..5658
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5707..5723)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5731..5747
                     /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(5755..5785)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5800..5821
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6109..6697)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6868..7728)
                     /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(7729..7833)
                     /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 ccaccatgaa gaggagtgaa
      721 aagccagaag gatatagaca aatgaggcct aagacctttc ctgccagtaa ctatactgtc
      781 agtagccggc aaatgttaca agaaattcgg gaatccctta ggaatttatc taaaccatct
      841 gatgctgcta aggctgagca taacatgagt aaaatgtcaa ccgaagatcc tcgacaagtc
      901 agaaatccac ccaaatttgg gacgcatcat aaagccttgc aggaaattcg aaactctctg
      961 cttccatttg caaatgaaac aaattcttct cggagtactt cagaagttaa tccacaaatg
     1021 cttcaagact tgcaagctgc tggatttgat gaggatatgg ttatacaagc tcttcagaaa
     1081 actaacaaca gaagtataga agcagcaatt gaattcatta gtaaaatgag ttaccaagat
     1141 cctcgacgag agcagatggc tgcagcagct gccagaccta ttaatgccag catgaaacca
     1201 gggaatgtgc agcaatcagt taaccgcaaa cagagctgga aaggttctaa agaatcctta
     1261 gttcctcaga ggcatggccc gccactagga gaaagtgtgg cctatcattc tgagagtccc
     1321 aactcacaga cagatgtagg aagacctttg tctggatctg gtatatcagc atttgttcaa
     1381 gctcacccta gcaacggaca gagagtgaac cccccaccac cacctcaagt aaggagtgtt
     1441 actcctccac cacctccaag aggccagact ccccctccaa gaggtacaac tccacctccc
     1501 ccttcatggg aaccaaactc tcaaacaaag cgctattctg gaaacatgga atacgtaatc
     1561 tcccgaatct ctcctgtccc acctggggca tggcaagagg gctatcctcc accacctctc
     1621 aacacttccc ccatgaatcc tcctaatcaa ggacagagag gcattagttc tgttcctgtt
     1681 ggcagacaac caatcatcat gcagagttct agcaaattta actttccatc agggagacct
     1741 ggaatgcaga atggtactgg acaaactgat ttcatgatac accaaaatgt tgtccctgct
     1801 ggcactgtga atcggcagcc accacctcca tatcctctga cagcagctaa tggacaaagc
     1861 ccttctgctt tacaaacagg gggatctgct gctccttcgt catatacaaa tggaagtatt
     1921 cctcagtcta tgatggtgcc aaacagaaat agtcataaca tggaactata taacattagt
     1981 gtacctggac tgcaaacaaa ttggcctcag tcatcttctg ctccagccca gtcatccccg
     2041 agcagtgggc atgaaatccc tacatggcaa cctaacatac cagtgaggtc aaattctttt
     2101 aataacccat taggaaatag agcaagtcac tctgctaatt ctcagccttc tgctacaaca
     2161 gtcactgcaa ttacaccagc tcctattcaa cagcctgtga aaagtatgcg tgtattaaaa
     2221 ccagagctac agactgcttt agcacctaca cacccttctt ggataccaca gccaattcaa
     2281 actgttcaac ccagtccttt tcctgaggga accgcttcaa atgtgactgt gatgccacct
     2341 gttgctgaag ctccaaacta tcaaggacca ccaccaccct acccaaaaca tctgctgcac
     2401 caaaacccat ctgttcctcc atacgagtca atcagtaagc ctagcaaaga ggatcagcca
     2461 agcttgccca aggaagatga gagtgaaaag agttatgaaa atgttgatag tggggataaa
     2521 gaaaagaaac agattacaac ttcacctatt actgttagga aaaacaagaa agatgaagag
     2581 cgaagggaat ctcgtattca aagttattct cctcaagcat ttaaattctt tatggagcaa
     2641 catgtagaaa atgtactcaa atctcatcag cagcgtctac atcgtaaaaa acaattagag
     2701 aatgaaatga tgcgggtaaa accttttaaa atgtccattt ttatacttaa tcatctgttt
     2761 gcttggtgtt tatttctcga gcaccatcac catcaccatc accattaggg cccgatatct
     2821 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2881 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2941 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3001 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3061 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3121 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3181 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3241 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3301 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3361 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     3421 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3481 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     3541 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3601 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3661 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3721 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3781 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4021 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4081 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4141 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4501 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4561 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4621 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4681 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4741 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4801 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4861 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4921 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4981 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5041 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5101 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5161 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5221 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5281 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5341 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     5401 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5461 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5521 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5581 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5641 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5701 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5761 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5821 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5881 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5941 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6301 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6541 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7501 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7561 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7621 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7681 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7801 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7861 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7921 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7981 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     8041 ctgcttagg
//
