LOCUS       sequence-316187-        3863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac ITR's from PB-CA retrofitted with a linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3863)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3863)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PB-CA retrofitted
FEATURES             Location/Qualifiers
     source          1..3863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   86..148
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   870..904
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(1053..1071)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1092..1108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1092..1108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1105..1127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1116..1132
                     /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(1140..1146,1147..1164,1165..1170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1185..1206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1323..1340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1494..2082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1574..1593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2253..3044,3045..3113))
                     /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     2876..2895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3114..3218)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3244..3699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3402)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3593..3612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3826..3848
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3840..3857
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3841..3857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctcggtattc acgacagcag gctgaataat
       61 aaaaaaatta gaaactatta tttaacccta gaaagataat catattgtga cgtacgttaa
      121 agataatcat gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
      181 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
      241 aacaaacaat ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct
      301 tctataaagt aacaaaactt ttaaacattc tctcttttac aaaaataaac ttattttgta
      361 ctttaaaaac agtcatgttg tattataaaa taagtaatta gcttaaccta tacataatag
      421 aaacaaatta tacttattag tcagtcagaa acaactttgg cacatatcaa tattatgctc
      481 tcgttaatac cggtcagtaa gcttctagcc aacaagctcg tcatcgcttt gcagaagagc
      541 agagaggata tgctcatcgt ctaaagaact acccatttta ttatatatta gtcacgatat
      601 ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat
      661 ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg
      721 actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc
      781 acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt
      841 tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct
      901 agggttaaaa aagatttgcg ctttactcga cctaaacttt aaacacgtca tagaatcttc
      961 gtttgacaaa aaccacattg tggccaagct gtgtgacgcg acgcgcgcta aagaatggca
     1021 aaccaagtcg cgcgaggtac cagcttttgt tccctttagt gagggttaat ttcgagcttg
     1081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3241 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3361 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     3721 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     3781 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     3841 gtaaaacgac ggccagtgaa ttg
//
