LOCUS       sequence-131712-        4410 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 4410)
  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 4410)
  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/SB10-Delta-DDE
FEATURES             Location/Qualifiers
     source          1..4410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..74
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     66..83
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     67..83
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        110..413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     568..588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     614..638
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          755..851
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     polyA_signal    1632..1766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1656..1675)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1710..1729
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1877..1893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1877..1893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1890..1912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1901..1917
                     /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(1925..1931,1932..1949,1950..1955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1970..1991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2108..2125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2279..2867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2359..2378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3038..3829,3830..3898))
                     /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     3661..3680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3899..4003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4071..4089
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4127..4149)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4249..4268
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
       61 gacgttgtaa aacgacggcc agtgaattcg agcttgcatg cctgcaggtc gttacataac
      121 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      181 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      241 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      301 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      361 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      421 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      481 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      541 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      601 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
      661 gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta gaggatccgg
      721 tactcgagga actgaaaaac cagaaagtta actggtaagt ttagtctttt tgtcttttat
      781 ttcaggtccc ggatccggtg gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc
      841 tttacttcta ggcctgtacg gaagtgttac ttctgctcta aaagctgcgg aattgtaccc
      901 gcggccgatc cgacatcatg ggaaaatcaa aagaaatcag ccaagacctc agaaaaaaaa
      961 ttgtagacct ccacaagtct ggttcatcct tgggagcaat ttccaaacgc ctgaaagtac
     1021 cacgttcatc tgtacaaaca atagtacgca agtataaaca ccatgggacc acgcagccgt
     1081 cataccgctc aggaaggaga cgcgttctgt ctcctagaga tgaacgtact ttggtgcgaa
     1141 aagtgcaaat caatcccaga acaacagcaa aggaccttgt gaagatgctg gaggaaacag
     1201 gtacaaaagt atctatatcc acagtaaaac gagtcctata tcgacataac ctgaaaggcc
     1261 gctcagcaag gaagaagcca ctgctccaaa accgacataa gaaagccaga ctacggtttg
     1321 caactgcaca tggggacaaa gatcgtactt tttggagaaa tgtcctctgg tctgatgaaa
     1381 caaaaataga actgtttggc catcacaaag ccctgacctc aatcctatag aaaatttgtg
     1441 ggcagaactg aaaaagcgtg tgcgagcaag gaggcctaca aacctgactc agttacacca
     1501 gctctgtcag gaggaatggg ccaaaattca cccaacttat tgtgggaagc ttgtggaagg
     1561 ctacccgaaa cgtttgaccc aagttaaaca atttaaaggc aatgctacca aatactagaa
     1621 ttggccgcgg ggatccagac atgataagat acattgatga gtttggacaa accacaacta
     1681 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1741 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1801 ttcaggggga ggtgtgggag gttttttcgg atcctctaga gtcgacctgc aggcatgcaa
     1861 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1921 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1981 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2041 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2101 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2461 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2701 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4021 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4081 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4141 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4201 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4261 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4321 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4381 tgcgggcctc ttcgctatta cgccagctgg
//
