LOCUS       sequence-372403-        3095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-3 vector for stable integration of up to three cassettes via 
            piggyBac transposase (ITR-A1::A2::A3-ITR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3095)
  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 3095)
  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: pTS1019_Tier3(PB)
FEATURES             Location/Qualifiers
     source          1..3095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     194..213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(432..536)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   596..658
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(952..969)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    958..1182
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1328..1344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1328..1344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1500..1521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   1851..1885
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(1957..2545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2037..2056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2666..3095,1..362,363..431))
                     /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 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
       61 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      121 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      181 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      241 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      301 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      361 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      421 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      481 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      541 tttccccgaa aagtgccacc tgacgtctaa gaaaccaaga tcagcggccg cttaacccta
      601 gaaagataat catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg
      661 ttttatcggt ctgtatatcg aggtttattt attaatttga atagatatta agttttatta
      721 tatttacact tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt
      781 attaaaaaaa aacaaaaact caaaatttct tctataaagt aacaaaacct taagagatct
      841 acgcgtggta ccctcgagga gctcttcgaa gcggaattca ccatgactag tgctagcgga
      901 tccaccggtg tctagaaagc tttgaggccg gcctgcaggc ccgctgatca gcctcgactg
      961 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1021 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1081 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1141 aagacaatag caggcatgct ggggatgcgg tgggctctat ggtagcttag gaggcggaaa
     1201 gaaccagcgc gggagaccgt cagttggtct ccagctatac aggtgcaccg ctttataata
     1261 aaactgatac agagaacgtt atcatgtgtg ttggtttttt ttgtgtgtgc ggtttaaggg
     1321 agcctcggtc atagctgttt cctgtgtaaa acgacggcca gcgcgggaga cggtcagttc
     1381 gtctccagct cagaaattat gagagcgttt aataaagtat tgactatggt tgacaaaacg
     1441 tgtgttggag ttttttgtgt gctttgtcat caactgagcc agtaggtcga gggtgcctaa
     1501 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1561 ctgtcgtgcc agctgcgata tctataacaa gaaaatatat atataataag ttatcacgta
     1621 agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa
     1681 agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta
     1741 ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca
     1801 cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa
     1861 ttttacgcag actatctttc tagggttaac catggacatg tgagcaaaag gccagcaaaa
     1921 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1981 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2041 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2101 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2161 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2221 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2281 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2341 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2401 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2461 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2521 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggattatcaa
     2581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcggctc ccacgctcac
     2821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3061 gatcccccat gttgtgcaaa aaagcggtta gctcc
//
