LOCUS       sequence-204450-        3457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized Sleeping Beauty Transposon Vector with MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3457)
  AUTHORS   Wang Y, Pryputniewicz-Dobrinska D, Nagy EE, Kaufman CD, Singh M, 
            Yant S, Wang J, Dalda A, Kay MA, Ivics Z, Izsvak Z
  TITLE     Regulated complex assembly safeguards the fidelity of Sleeping 
            Beauty transposition.
  JOURNAL   Nucleic Acids Res. 2017 Jan 9;45(1):311-326. doi: 
            10.1093/nar/gkw1164. Epub 2016 Dec 1.
  PUBMED    27913727
REFERENCE   2  (bases 1 to 3457)
  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"; doi: 
            "10.1093/nar/gkw1164"; journalName: "Nucleic Acids Res"; date: 
            "2017-01-9- 9"; volume: "45"; issue: "1"; pages: "311-326"
COMMENT     Sequence Label: pT4/HB
FEATURES             Location/Qualifiers
     source          1..3457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..56
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     48..65
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     49..65
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..94
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        75..93
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(718..738)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(718..736)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(757..773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(757..773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(770..792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    781..797
                     /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(805..811,812..829,830..835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    850..871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(988..1005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1159..1747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1239..1258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1918..2709,2710..2778))
                     /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     2541..2560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2779..2883)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2910..3365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2997..3016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3207..3228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
       61 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctagcg tttaaaccta
      121 cagttgaagt cggaagttta catacactta agttggagtc attaaaactc gtttttcaac
      181 tactccacaa atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact
      241 ttgtgcatga cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata
      301 attcactgta tcacaattcc agtgggtcag aagtgtacat acacgcgctt gactgtgcct
      361 ttaagcttga tatccatgga attcactagt gcgcgcggcc gctctagatg gccagatctt
      421 taaacaattt aaaggcaatg ctaccaaata ctaagcgcgt gtatgtacac ttctgaccca
      481 ctgggaatgt gatgaaagaa ataaaagctg aaatgaatca ttctctctac tattattctg
      541 atatttcaca ttcttaaaat aaagtggtga tcctaactga ccttaagaca gggaatcttt
      601 actcggatta aatgtcagga attgtgaaaa agtgagttta aatgtatttg gctaaggtgt
      661 atgtaaactt ccgacttcaa ctgtagttta aacgggcccg gtacccagct tttgttccct
      721 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      781 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      841 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      901 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      961 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1021 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1561 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1861 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1921 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1981 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2041 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2101 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2161 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2221 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2281 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2341 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2401 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2461 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2521 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2581 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2641 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2701 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2761 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2821 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2881 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     2941 gggagtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3001 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3061 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3121 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3181 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3241 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3301 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3361 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     3421 tcgctattac gccagctggc gaaaggggga tgtgctg
//
