LOCUS       V033246                 9767 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033246
VERSION     V033246
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             896..2965
                     /label="LATS1(NM_001270519)"
                     /note="LATS1(NM_001270519)"
                     /gene="LATS1"
     CDS             2981..3037
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3047..3763
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced CFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG
                     IKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="ECFP"
     polyA_signal    5334..5558
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5604..6032
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6046..6375
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6442..7236
                     /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    7410..7531
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7580..7596)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7604..7620
                     /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(7628..7658)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7673..7694
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7982..8567)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8738..9598)
                     /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(9599..9703)
                     /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 ccaccatgaa
      901 gaggagtgaa aagccagaag gatatagaca aatgaggcct aagacctttc ctgccagtaa
      961 ctatactgtc agtagccggc aaatgttaca agaaattcgg gaatccctta ggaatttatc
     1021 taaaccatct gatgctgcta aggctgagca taacatgagt aaaatgtcaa ccgaagatcc
     1081 tcgacaagtc agaaatccac ccaaatttgg gacgcatcat aaagccttgc aggaaattcg
     1141 aaactctctg cttccatttg caaatgaaac aaattcttct cggagtactt cagaagttaa
     1201 tccacaaatg cttcaagact tgcaagctgc tggatttgat gaggatatgg ttatacaagc
     1261 tcttcagaaa actaacaaca gaagtataga agcagcaatt gaattcatta gtaaaatgag
     1321 ttaccaagat cctcgacgag agcagatggc tgcagcagct gccagaccta ttaatgccag
     1381 catgaaacca gggaatgtgc agcaatcagt taaccgcaaa cagagctgga aaggttctaa
     1441 agaatcctta gttcctcaga ggcatggccc gccactagga gaaagtgtgg cctatcattc
     1501 tgagagtccc aactcacaga cagatgtagg aagacctttg tctggatctg gtatatcagc
     1561 atttgttcaa gctcacccta gcaacggaca gagagtgaac cccccaccac cacctcaagt
     1621 aaggagtgtt actcctccac cacctccaag aggccagact ccccctccaa gaggtacaac
     1681 tccacctccc ccttcatggg aaccaaactc tcaaacaaag cgctattctg gaaacatgga
     1741 atacgtaatc tcccgaatct ctcctgtccc acctggggca tggcaagagg gctatcctcc
     1801 accacctctc aacacttccc ccatgaatcc tcctaatcaa ggacagagag gcattagttc
     1861 tgttcctgtt ggcagacaac caatcatcat gcagagttct agcaaattta actttccatc
     1921 agggagacct ggaatgcaga atggtactgg acaaactgat ttcatgatac accaaaatgt
     1981 tgtccctgct ggcactgtga atcggcagcc accacctcca tatcctctga cagcagctaa
     2041 tggacaaagc ccttctgctt tacaaacagg gggatctgct gctccttcgt catatacaaa
     2101 tggaagtatt cctcagtcta tgatggtgcc aaacagaaat agtcataaca tggaactata
     2161 taacattagt gtacctggac tgcaaacaaa ttggcctcag tcatcttctg ctccagccca
     2221 gtcatccccg agcagtgggc atgaaatccc tacatggcaa cctaacatac cagtgaggtc
     2281 aaattctttt aataacccat taggaaatag agcaagtcac tctgctaatt ctcagccttc
     2341 tgctacaaca gtcactgcaa ttacaccagc tcctattcaa cagcctgtga aaagtatgcg
     2401 tgtattaaaa ccagagctac agactgcttt agcacctaca cacccttctt ggataccaca
     2461 gccaattcaa actgttcaac ccagtccttt tcctgaggga accgcttcaa atgtgactgt
     2521 gatgccacct gttgctgaag ctccaaacta tcaaggacca ccaccaccct acccaaaaca
     2581 tctgctgcac caaaacccat ctgttcctcc atacgagtca atcagtaagc ctagcaaaga
     2641 ggatcagcca agcttgccca aggaagatga gagtgaaaag agttatgaaa atgttgatag
     2701 tggggataaa gaaaagaaac agattacaac ttcacctatt actgttagga aaaacaagaa
     2761 agatgaagag cgaagggaat ctcgtattca aagttattct cctcaagcat ttaaattctt
     2821 tatggagcaa catgtagaaa atgtactcaa atctcatcag cagcgtctac atcgtaaaaa
     2881 acaattagag aatgaaatga tgcgggtaaa accttttaaa atgtccattt ttatacttaa
     2941 tcatctgttt gcttggtgtt tatttggatc cggaagcgga gccactaact tctccctgtt
     3001 gaaacaagca ggggatgtcg aagagaatcc cgggccaggt ggatcaatgg tgagcaaggg
     3061 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3121 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3181 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3241 gacctggggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3301 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3361 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3421 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3481 ctacatcagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     3541 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3601 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3661 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3721 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggaattca tggatcccgg
     3781 gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt cgcagccccc
     3841 gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga cccaggcggc
     3901 gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact cggagaccga
     3961 cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc cccagaccgt
     4021 gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc ccaaatccca
     4081 ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac agcatgttcg
     4141 agctcattcc tctccagctt ctctgcagtt gggagctgtt tctcctggga cactgacccc
     4201 cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc gacagtcttc
     4261 ttttgagata cctgatgatg tacctctgcc agcaggttgg gagatggcaa agacatcttc
     4321 tggtcagaga tacttcttaa atcacatcga tcagacaaca acatggcagg accccaggaa
     4381 ggccatgctg tcccagatga acgtcacagc ccccaccagt ccaccagtgc agcagaatat
     4441 gatgaactcg gcttcaggtc ctcttcctga tggatgggaa caagccatga ctcaggatgg
     4501 agaaatttac tatataaacc ataagaacaa gaccacctct tggctagacc caaggcttga
     4561 ccctcgtttt gccatgaacc agagaatcag tcagagtgct ccagtgaaac agccaccacc
     4621 cctggctccc cagagcccac agggaggcgt catgggtggc agcaactcca accagcagca
     4681 acagatgcga ctgcagcaac tgcagatgga gaaggagagg ctgcggctga aacagcaaga
     4741 actgcttcgg caggcaatgc ggaatatcaa tcccagcaca gcaaattctc caaaatgtca
     4801 ggagttagcc ctgcgtagcc agttaccaac actggagcag gatggtggga ctcaaaatcc
     4861 agtgtcttct cccgggatgt ctcaggaatt gagaacaatg acgaccaata gctcagatcc
     4921 tttccttaac agtggcacct atcactctcg agatgagagt acagacagtg gactaagcat
     4981 gagcagctac agtgtccctc gaaccccaga tgacttcctg aacagtgtgg atgagatgga
     5041 tacaggtgat actatcaacc aaagcaccct gccctcacag cagaaccgtt tcccagacta
     5101 ccttgaagcc attcctggga caaatgtgga ccttggaaca ctggaaggag atggaatgaa
     5161 catagaagga gaggagctga tgccaagtct gcaggaagct ttgagttctg acatccttaa
     5221 tgacatggag tctgttttgg ctgccaccaa gctagataaa gaaagctttc ttacatggtt
     5281 aaccggttag ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     5341 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5401 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5461 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5521 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     5581 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5941 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6001 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     6061 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6121 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6181 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6241 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6301 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6361 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     6421 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     6481 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     6541 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     6601 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     6661 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     6721 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     6781 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     6841 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6901 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6961 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     7021 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     7081 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     7141 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     7201 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     7261 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     7321 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     7381 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     7441 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7501 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     7561 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7621 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7681 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7741 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7801 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7861 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7921 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7981 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     8041 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     8101 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     8161 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     8221 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     8281 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     8341 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     8401 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     8461 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     8521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     9001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     9061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9721 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
