LOCUS       sequence-365642-        3108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-3 vector for stable integration of up to three cassettes via 
            sleeping beauty transposase (5'ITR-A1-pA::A2-pA::A3-pA-3'ITR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3108)
  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 3108)
  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: pTS1107_Tier3(SB)
FEATURES             Location/Qualifiers
     source          1..3108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(230..1021,1022..1090))
                     /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     853..872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1091..1195)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1658..1675)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1664..1888
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2034..2050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2034..2050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2206..2227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(join(2629..3108,1..109))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2709..2728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
       61 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      121 gatcttttct acggggatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      181 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      241 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      301 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
      361 tgataccgcg gctcccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
      421 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
      481 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
      541 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
      601 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
      661 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
      721 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
      781 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
      841 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
      901 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      961 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1021 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1081 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1141 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1201 ttccccgaaa agtgccacct gacgtctaag aaaccaagat cagcggccgc tacagttgaa
     1261 gtcggaagtt tacatacagt taagttggag tcattaaaac tcgtttttca actactccac
     1321 aaatttcttg ttaacaaaca atagttttgg caagtcagtt aggacatcta ctttgtgcat
     1381 gacacaagtc atttttccaa caattgttta cagacagatt atttcactta taattcactg
     1441 tatcacaatt ccagtgggtc agaagtttac atacactaag ttcgactcct ctgcagaatg
     1501 cggcgatgtt tcggtaaggg gtccgctatc tagtcttaag agatctacgc gtggtaccct
     1561 cgaggagctc ttcgaagcgg aattcaccat gactagtgct agcggatcca ccggtgtcta
     1621 gaaagctttg aggccggcct gcaggcccgc tgatcagcct cgactgtgcc ttctagttgc
     1681 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1741 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     1801 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     1861 catgctgggg atgcggtggg ctctatggta gcttaggagg cggaaagaac cagcgcggga
     1921 gaccgtcagt tggtctccag ctatacaggt gcaccgcttt ataataaaac tgatacagag
     1981 aacgttatca tgtgtgttgg ttttttttgt gtgtgcggtt taagggagcc tcggtcatag
     2041 ctgtttcctg tgtaaaacga cggccagcgc gggagacggt cagttcgtct ccagctcaga
     2101 aattatgaga gcgtttaata aagtattgac tatggttgac aaaacgtgtg ttggagtttt
     2161 ttgtgtgctt tgtcatcaac tgagccagta ggtcgagggt gcctaatgag tgagctaact
     2221 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2281 gctagctact cgggacccct taccgaaaca tcgccgcatt ctgcagagga gtcgagtgta
     2341 tgtaaacttc tgacccactg ggaatgtgat gaaagaaata aaagctgaaa tgaatcattc
     2401 tctctactat tattctgata tttcacattc ttaaaataaa gtggtgatcc taactgacct
     2461 aagacaggga atttttacta ggattaaatg tcaggaattg tgaaaaagtg agtttaaatg
     2521 tatttggcta aggtgtatgt aaacttccga cttcaactgt accatggaca tgtgagcaaa
     2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttac
//
