LOCUS       sequence-12571-e        4732 bp    DNA     circular SYN 15-DEC-2025
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1997 
            Nov 14. 91(4):501-10."
COMMENT     Sequence Label: pCMV/SB11
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..328
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        329..532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     483..503
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     529..553
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          670..766
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             833..1855
                     /codon_start=1
                     /product="hyperactive variant of the Sleeping Beauty 
                     transposase (Mates et al., 2009)"
                     /label=SB100X
                     /translation="MGKSKEISQDLRKKIVDLHKSGSSLGAISKRLKVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGRSARKKPLLQNRHKKARLRFARAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMRKENYVDIL
                     KQHLKTSVRKLKLGRKWVFQQDNDPKHTSKHVRKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPTYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     polyA_signal    1869..2003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1893..1912)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1947..1966
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2114..2130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2114..2130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2127..2149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2138..2154
                     /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(join(2162..2168,2169..2186,2187..2192))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2207..2228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2345..2362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2516..3104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2596..2615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3275..4066,4067..4135))
                     /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     3898..3917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4136..4240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4308..4326
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4364..4386)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4486..4505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4699..4721
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4713..4730
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4714..4730
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc cgcctggctg
       61 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      121 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      181 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      541 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
      601 ggaccgatcc agcctccgga ctctagagga tccggtactc gaggaactga aaaaccagaa
      661 agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt
      721 gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt
      781 gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgatccgaca tcatgggaaa
      841 atcaaaagaa atcagccaag acctcagaaa aaaaattgta gacctccaca agtctggttc
      901 atccttggga gcaatttcca aacgcctgaa agtaccacgt tcatctgtac aaacaatagt
      961 acgcaagtat aaacaccatg ggaccacgca gccgtcatac cgctcaggaa ggagacgcgt
     1021 tctgtctcct agagatgaac gtactttggt gcgaaaagtg caaatcaatc ccagaacaac
     1081 agcaaaggac cttgtgaaga tgctggagga aacaggtaca aaagtatcta tatccacagt
     1141 aaaacgagtc ctatatcgac ataacctgaa aggccgctca gcaaggaaga agccactgct
     1201 ccaaaaccga cataagaaag ccagactacg gtttgcaaga gcacatgggg acaaagatcg
     1261 tactttttgg agaaatgtcc tctggtctga tgaaacaaaa atagaactgt ttggccataa
     1321 tgaccatcgt tatgtttgga ggaagaaggg ggaggcttgc aagccgaaga acaccatccc
     1381 aaccgtgaag cacgggggtg gcagcatcat gttgtggggg tgctttgctg caggagggac
     1441 tggtgcactt cacaaaatag atggcatcat gaggaaggaa aattatgtgg atatattgaa
     1501 gcaacatctc aagacatcag tcaggaagtt aaagcttggt cgcaaatggg tcttccaaca
     1561 agacaatgac cccaagcata cttccaaaca cgtgagaaaa tggcttaagg acaacaaagt
     1621 caaggtattg gagtggccat cacaaagccc tgacctcaat cctatagaaa atttgtgggc
     1681 agaactgaaa aagcgtgtgc gagcaaggag gcctacaaac ctgactcagt tacaccagct
     1741 ctgtcaggag gaatgggcca aaattcaccc aacttattgt gggaagcttg tggaaggcta
     1801 cccgaaacgt ttgacccaag ttaaacaatt taaaggcaat gctaccaaat actagaattg
     1861 gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1921 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1981 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2041 agggggaggt gtgggaggtt ttttcggatc ctctagagtc gacctgcagg catgcaagct
     2101 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2161 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2221 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2281 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2941 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4201 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4261 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4321 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4381 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4441 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4501 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4561 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4621 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4681 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt ga
//
