LOCUS       sequence-366198-        4153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-3 vector for stable stable integration of up to three cassettes
            via sleeping beauty transposase including a ZeoR resistance gene 
            (5'ITR-A1-pA::A2-pA::PRPBSA-ZeoR-pA-3'ITR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4153)
  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 4153)
  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: pTS2351_Tier3(SB)-Zeo
FEATURES             Location/Qualifiers
     source          1..4153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     621..640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(859..963)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1426..1443)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1432..1656
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1802..1818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1802..1818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2508..2879
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     protein_bind    3019..3040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3442..4030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3522..3541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4151..4153,1..789,790..858))
                     /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"
ORIGIN
        1 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
       61 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      121 tgctgcaatg ataccgcggc tcccacgctc accggctcca gatttatcag caataaacca
      181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      961 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accaagatca gcggccgcta
     1021 cagttgaagt cggaagttta catacagtta agttggagtc attaaaactc gtttttcaac
     1081 tactccacaa atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact
     1141 ttgtgcatga cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata
     1201 attcactgta tcacaattcc agtgggtcag aagtttacat acactaagtt cgactcctct
     1261 gcagaatgcg gcgatgtttc ggtaaggggt ccgctatcta gtcttaagag atctacgcgt
     1321 ggtaccctcg aggagctctt cgaagcggaa ttcaccatga ctagtgctag cggatccacc
     1381 ggtgtctaga aagctttgag gccggcctgc aggcccgctg atcagcctcg actgtgcctt
     1441 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1501 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1561 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1621 atagcaggca tgctggggat gcggtgggct ctatggtagc ttaggaggcg gaaagaacca
     1681 gcgcgggaga ccgtcagttg gtctccagct atacaggtgc accgctttat aataaaactg
     1741 atacagagaa cgttatcatg tgtgttggtt ttttttgtgt gtgcggttta agggagcctc
     1801 ggtcatagct gtttcctgtg taaaacgacg gccagcgcgt ggtaccctcg agcgtgaccc
     1861 tgtctcacaa aataaagtaa gcccggactg agtgcggaaa ggcgggcctg gcgggtctgg
     1921 actccccatg cgggccacca gaggccctgc agccttcagt cgcttgaagg ggtaatggcg
     1981 cttccactca caaacatggc ggacagagcg tgtgaacgag atgaacagcc cctcaaaaat
     2041 atggccgccg aggctggacg gccgtgcccc agcagcaccg cctccgcgcc ccacgtgatc
     2101 tctcgccggg cacagcgctg accgcggagg tccaaccgga agaatgtccg gattggacat
     2161 tcggaagagg gcccgccttc cctggggaat ctctgcgcac gcgcagaacg cttcgaccaa
     2221 tgaaaacaca ggaagccgtc cgcgcaaccg cgttgcgtca cttctgccgc ccctgtttca
     2281 aggtatatag ccgtagacgg aacttcgcct ttctctcggc cttagcgcca tttttttggg
     2341 tgagtgtttt ttggttcctg cgttgggatt ccgtgtacaa tccatagaca tctgacctcg
     2401 gcacttagca tcatcacagc aaactaactg tagcctttct ctctttccct gtagaaacct
     2461 ctgcacctag ggagctcgaa ttcaccatga ctagtggtgg ttctggtatg gccaagttga
     2521 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     2581 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     2641 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     2701 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     2761 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     2821 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggacg
     2881 ctagcggatc caccggtgtc tagaaagctc agaaattatg agagcgttta ataaagtatt
     2941 gactatggtt gacaaaacgt gtgttggagt tttttgtgtg ctttgtcatc aactgagcca
     3001 gtaggtcgag ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3061 cgctttccag tcgggaaacc tgtcgtgcca gctgctagct actcgggacc ccttaccgaa
     3121 acatcgccgc attctgcaga ggagtcgagt gtatgtaaac ttctgaccca ctgggaatgt
     3181 gatgaaagaa ataaaagctg aaatgaatca ttctctctac tattattctg atatttcaca
     3241 ttcttaaaat aaagtggtga tcctaactga cctaagacag ggaattttta ctaggattaa
     3301 atgtcaggaa ttgtgaaaaa gtgagtttaa atgtatttgg ctaaggtgta tgtaaacttc
     3361 cgacttcaac tgtaccatgg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3841 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4021 aggatctcaa gaagatcctt tgatcttttc tacggggatt atcaaaaagg atcttcacct
     4081 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4141 ggtctgacag tta
//
