LOCUS       Exported                3237 bp DNA     circular SYN 03-DEC-2024
DEFINITION  To all for PiggyBac gene insertion into Genome.
ACCESSION   .
VERSION     .
KEYWORDS    PiggyBac PlayBack
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3237)
  AUTHORS   Foran G, Hallam RD, Megaly M, Turgambayeva A, Necakov A
  TITLE     PlayBack cloning: simple, reversible, cost-effective cloning for the
            combinatorial assembly of complex expression constructs.
  JOURNAL   Biotechniques. 2023 Oct;75(4):168-178. doi: 10.2144/btn-2023-0042. 
            Epub 2023 Oct 10.
  PUBMED    37815818
REFERENCE   2  (bases 1 to 3237)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 3, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.2144/btn-2023-0042"; journalName: "Biotechniques"; date: 
            "2023-10"; volume: "75"; issue: "4"; pages: "168-178"
FEATURES             Location/Qualifiers
     source          1..3237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     repeat_region   194..228
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal
                     repeat sequence (ITR)"
     repeat_region   931..993
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal
                     repeat sequence (ITR)"
     primer_bind     complement(1016..1032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1016..1032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1029..1051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1040..1056
                     /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(1064..1094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1109..1130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1418..2006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1498..1517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2177..3037)
                     /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     2800..2819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3038..3142)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3210..3228
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgtt taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      241 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      301 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      361 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      421 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      481 atattttctt gttatagata tcgtcacgcg tagcgagtca gtgagcgagg aagcggaaga
      541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      601 cctcgaggac aggtttcccg actggaaagc gggcagcgag tcagtgagcg aggaagcgga
      661 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      721 gcacgacagg tttcccgact ggaaaggtcg accggggaaa gttttgttac tttatagaag
      781 aaattttgag tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga
      841 atttattatt agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa
      901 ataaacctcg atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt
      961 aacgtacgtc acaatatgat tatctttcta gggttaaaag cttggcgtaa tcatggtcat
     1021 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1081 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1141 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1201 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1261 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1321 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2461 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2581 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2701 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2761 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2821 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2881 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2941 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3001 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3061 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3121 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3181 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
