LOCUS       sequence-274419-        3680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac ITR's in high copy number pBluescript backbone plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3680)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3680)
  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            "Unpublished"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: pBluescript-PB
FEATURES             Location/Qualifiers
     source          1..3680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..96
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     88..105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     89..105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     112..131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        112..130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   218..252
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   854..916
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(980..1000)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(980..998)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1019..1035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1019..1035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1032..1054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1043..1059
                     /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(1067..1073,1074..1091,1092..1097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1112..1133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1250..1267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1421..2009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1501..1520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2180..2971,2972..3040))
                     /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     2803..2822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3041..3145)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3172..3627
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3259..3278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3469..3490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
       61 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
      121 tcactatagg gcgacccaac acaatgtggt ttttgtcaaa cgaagattct atgacgtgtt
      181 taaagtttag gtcgagtaaa gcgcaaatct tttttaaccc tagaaagata gtctgcgtaa
      241 aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg aatccgtcgc
      301 tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc ttgtcaatgc
      361 ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac gcatgattat
      421 cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt tcatgttcta
      481 cttacgtgat aacttattat atatatattt tcttgttata gatatcgtga ctaatatata
      541 ataaaaaagc ttactgaccg gttaataagt ataatttgtt tctattatgt ataggttaag
      601 ctaattactt attttataat acaacatgac tgtttttaaa gtacaaaata agtttatttt
      661 tgtaaaagag agaatgttta aaagttttgt tactttatag aagaaatttt gagtttttgt
      721 ttttttttaa taaataaata aacataaata aattgtttgt tgaatttatt attagtatgt
      781 aagtgtaaat ataataaaac ttaatatcta ttcaaattaa taaataaacc tcgatataca
      841 gaccgataaa acacatgcgt caattttacg catgattatc tttaacgtac gtcacaatat
      901 gattatcttt ctagggttaa ataatagttt ctaatttttt tattattcag cctgctgtcg
      961 tgaataccgg gggaggttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     1021 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1081 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1141 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1201 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1261 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1321 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2161 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2341 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2401 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2461 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2581 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2701 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2821 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2881 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3061 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3121 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     3181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3241 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3361 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3481 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3541 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3601 ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg
     3661 aagggcgatc ggtgcgggcc
//
