LOCUS        pCMV/SB10        4732 bp DNA     circular SYN 25-AUG-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  AUTHORS   Ivics Z, Hackett PB, Plasterk RH, Izsvak Z
  TITLE     Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon 
            from fish, and its transposition in human cells.
  JOURNAL   Cell. 1997 Nov 14. 91(4):501-10.
  PUBMED    9390559
REFERENCE   2  (bases 1 to 4732)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2024 from SnapGene Server 7.0.3
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1997 
            Nov 14. 91(4):501-10."
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(953..1813)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1986..2004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2042..2064)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2164..2183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2377..2399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2391..2408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2392..2408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2435..2738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2739..2942
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2893..2913
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2939..2963
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3080..3176
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             3243..4265
                     /codon_start=1
                     /product="hyperactive variant of the Sleeping Beauty 
                     transposase (Mates et al., 2009)"
                     /label=SB100X
                     /translation="MGKSKEISQDLRKKIVDLHKSGSSLGAISKRLKVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGRSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMRKENYVDIL
                     KQHLKTSVRKLKLGRKWVFQMDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPTYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     polyA_signal    4279..4413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4303..4322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4357..4376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4524..4540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4524..4540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4537..4559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4548..4564
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4572..4602)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4617..4638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1981 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2041 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2101 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     2161 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     2221 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     2281 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     2341 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     2401 cggccagtga attcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc
     2461 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2521 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2581 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2641 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2701 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2761 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2821 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2881 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2941 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     3001 gaagacaccg ggaccgatcc agcctccgga ctctagagga tccggtactc gaggaactga
     3061 aaaaccagaa agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc
     3121 cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct
     3181 gtacggaagt gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgatccgaca
     3241 tcatgggaaa atcaaaagaa atcagccaag acctcagaaa aaaaattgta gacctccaca
     3301 agtctggttc atccttggga gcaatttcca aacgcctgaa agtaccacgt tcatctgtac
     3361 aaacaatagt acgcaagtat aaacaccatg ggaccacgca gccgtcatac cgctcaggaa
     3421 ggagacgcgt tctgtctcct agagatgaac gtactttggt gcgaaaagtg caaatcaatc
     3481 ccagaacaac agcaaaggac cttgtgaaga tgctggagga aacaggtaca aaagtatcta
     3541 tatccacagt aaaacgagtc ctatatcgac ataacctgaa aggccgctca gcaaggaaga
     3601 agccactgct ccaaaaccga cataagaaag ccagactacg gtttgcaact gcacatgggg
     3661 acaaagatcg tactttttgg agaaatgtcc tctggtctga tgaaacaaaa atagaactgt
     3721 ttggccataa tgaccatcgt tatgtttgga ggaagaaggg ggaggcttgc aagccgaaga
     3781 acaccatccc aaccgtgaag cacgggggtg gcagcatcat gttgtggggg tgctttgctg
     3841 caggagggac tggtgcactt cacaaaatag atggcatcat gaggaaggaa aattatgtgg
     3901 atatattgaa gcaacatctc aagacatcag tcaggaagtt aaagcttggt cgcaaatggg
     3961 tcttccaaat ggacaatgac cccaagcata cttccaaagt tgtggcaaaa tggcttaagg
     4021 acaacaaagt caaggtattg gagtggccat cacaaagccc tgacctcaat cctatagaaa
     4081 atttgtgggc agaactgaaa aagcgtgtgc gagcaaggag gcctacaaac ctgactcagt
     4141 tacaccagct ctgtcaggag gaatgggcca aaattcaccc aacttattgt gggaagcttg
     4201 tggaaggcta cccgaaacgt ttgacccaag ttaaacaatt taaaggcaat gctaccaaat
     4261 actagaattg gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc
     4321 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     4381 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     4441 tttcaggttc agggggaggt gtgggaggtt ttttcggatc ctctagagtc gacctgcagg
     4501 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gc
//