LOCUS       sequence-204001-        4732 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     primer_bind     76..98
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     90..107
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     91..107
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        134..437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        438..641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     592..612
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..662
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          779..875
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             942..1964
                     /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    1978..2112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2002..2021)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2056..2075
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2223..2239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2223..2239)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2236..2258)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2247..2263
                     /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(2271..2277,2278..2295,2296..2301))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2316..2337
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2454..2471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2625..3213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2705..2724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3384..4175,4176..4244))
                     /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     4007..4026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4245..4349)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4417..4435
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4473..4495)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4595..4614
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
       61 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcgagcttg
      121 catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      181 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      241 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      301 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      361 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      421 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      481 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      541 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      601 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      661 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      721 gcctccggac tctagaggat ccggtactcg aggaactgaa aaaccagaaa gttaactggt
      781 aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg caaatcaaag
      841 aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg ttacttctgc
      901 tctaaaagct gcggaattgt acccgcggcc gatccgacat catgggaaaa tcaaaagaaa
      961 tcagccaaga cctcagaaaa aaaattgtag acctccacaa gtctggttca tccttgggag
     1021 caatttccaa acgcctgaaa gtaccacgtt catctgtaca aacaatagta cgcaagtata
     1081 aacaccatgg gaccacgcag ccgtcatacc gctcaggaag gagacgcgtt ctgtctccta
     1141 gagatgaacg tactttggtg cgaaaagtgc aaatcaatcc cagaacaaca gcaaaggacc
     1201 ttgtgaagat gctggaggaa acaggtacaa aagtatctat atccacagta aaacgagtcc
     1261 tatatcgaca taacctgaaa ggccgctcag caaggaagaa gccactgctc caaaaccgac
     1321 ataagaaagc cagactacgg tttgcaagag cacatgggga caaagatcgt actttttgga
     1381 gaaatgtcct ctggtctgat gaaacaaaaa tagaactgtt tggccataat gaccatcgtt
     1441 atgtttggag gaagaagggg gaggcttgca agccgaagaa caccatccca accgtgaagc
     1501 acgggggtgg cagcatcatg ttgtgggggt gctttgctgc aggagggact ggtgcacttc
     1561 acaaaataga tggcatcatg aggaaggaaa attatgtgga tatattgaag caacatctca
     1621 agacatcagt caggaagtta aagcttggtc gcaaatgggt cttccaacaa gacaatgacc
     1681 ccaagcatac ttccaaacac gtgagaaaat ggcttaagga caacaaagtc aaggtattgg
     1741 agtggccatc acaaagccct gacctcaatc ctatagaaaa tttgtgggca gaactgaaaa
     1801 agcgtgtgcg agcaaggagg cctacaaacc tgactcagtt acaccagctc tgtcaggagg
     1861 aatgggccaa aattcaccca acttattgtg ggaagcttgt ggaaggctac ccgaaacgtt
     1921 tgacccaagt taaacaattt aaaggcaatg ctaccaaata ctagaattgg ccgcggggat
     1981 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2041 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2101 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2161 tgggaggttt tttcggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca
     2221 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2281 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2341 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2401 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2461 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2521 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2581 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2641 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2701 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2761 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2821 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2881 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2941 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3001 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3061 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3121 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3181 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3241 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3301 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3361 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3421 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3481 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3541 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3601 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3661 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3721 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3781 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3841 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3901 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3961 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4021 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4081 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4141 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4201 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4261 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4321 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4381 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4441 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4501 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4561 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4621 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4681 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg gg
//
