LOCUS       sequence-365222-        4060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector encoding PhCMV driven SB100 sleeping beauty 
            transposase (PhCMV-SB100-pA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4060)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 4060)
  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: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pTS395_PhCMV-SB100
FEATURES             Location/Qualifiers
     source          1..4060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..464
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        465..668
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     619..639
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     713..732
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        713..731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             775..1797
                     /codon_start=1
                     /product="hyperactive variant of the Sleeping Beauty 
                     transposase (Mates et al., 2009)"
                     /label=SB100X
                     /translation="MGKSKEISQDLRKRIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGHSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMDAVQYVDIL
                     KQHLKTSVRKLKLGRKWVFQHDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPNYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     primer_bind     complement(1861..1878)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1867..2091
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2179..2196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2350..2938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2430..2449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3109..3900,3901..3969))
                     /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     3732..3751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(3970..4060,1..14))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
       61 ctccacgcgt ggtaccctcg agttgacatt gattattgag tagttattaa tagtaatcaa
      121 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      181 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      421 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      481 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      541 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      601 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      661 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
      721 ctcactatag ggagacccaa gctgttcgaa gcggaattca ccatgactag taccatggga
      781 aaatcaaaag aaatcagcca agacctcaga aaaagaattg tagacctcca caagtctggt
      841 tcatccttgg gagcaatttc caaacgcctg gcggtaccac gttcatctgt acaaacaata
      901 gtacgcaagt ataaacacca tgggaccacg cagccgtcat accgctcagg aaggagacgc
      961 gttctgtctc ctagagatga acgtactttg gtgcgaaaag tgcaaatcaa tcccagaaca
     1021 acagcaaagg accttgtgaa gatgctggag gaaacaggta caaaagtatc tatatccaca
     1081 gtaaaacgag tcctatatcg acataacctg aaaggccact cagcaaggaa gaagccactg
     1141 ctccaaaacc gacataagaa agccagacta cggtttgcaa ctgcacatgg ggacaaagat
     1201 cgtacttttt ggagaaatgt cctctggtct gatgaaacaa aaatagaact gtttggccat
     1261 aatgaccatc gttatgtttg gaggaagaag ggggaggctt gcaagccgaa gaacaccatc
     1321 ccaaccgtga agcacggggg tggcagcatc atgttgtggg ggtgctttgc tgcaggaggg
     1381 actggtgcac ttcacaaaat agatggcatc atggacgcgg tgcagtatgt ggatatattg
     1441 aagcaacatc tcaagacatc agtcaggaag ttaaagcttg gtcgcaaatg ggtcttccaa
     1501 cacgacaatg accccaagca tacttccaaa gttgtggcaa aatggcttaa ggacaacaaa
     1561 gtcaaggtat tggagtggcc atcacaaagc cctgacctca atcctataga aaatttgtgg
     1621 gcagaactga aaaagcgtgt gcgagcaagg aggcctacaa acctgactca gttacaccag
     1681 ctctgtcagg aggaatgggc caaaattcac ccaaattatt gtgggaagct tgtggaaggc
     1741 tacccgaaac gtttgaccca agttaaacaa tttaaaggca atgctaccaa atactagggg
     1801 ccctaaccgc ggggatccac cggtgtctag aaagctttga ggccggcctg cagggatcag
     1861 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1921 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1981 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2041 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2101 cggaaagaac cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2161 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2221 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2281 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2341 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2401 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2461 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2521 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2581 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2641 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2701 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2761 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2821 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2881 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2941 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3001 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3061 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3241 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3421 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3721 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3961 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4021 gagcggatac atatttgaat gtatttagaa aaataaacaa
//
