LOCUS       sequence-348859-        3071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minimal 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 3071)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3071)
  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            "Unpublished"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-minimal PB
FEATURES             Location/Qualifiers
     source          1..3071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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)."
     primer_bind     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     228..248
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        230..248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   270..332
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   417..451
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(488..507)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(489..507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(514..531)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(514..530)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(523..545)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(672..1127)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(809..830)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1021..1040
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1154..1258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1259..1327,1328..2119)
                     /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     complement(1477..1496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2290..2878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2779..2798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3032..3049
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      241 actaaaggga acctccccca attccttaac cctagaaaga taatcatatt gtgacgtacg
      301 ttaaagataa tcatgcgtaa aattgacgca tgaggtctga agaggagttt acgtccagcc
      361 aaaccggtca tggatccatg aagcttgctc gactagagct tgcggaaccc ttcgaacatg
      421 cgtcaatttt acgcagacta tctttctagg gttaaatcag tcaggtacat aatggatctt
      481 tgggtcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
      541 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      601 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      661 tggcgaatgg caaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
      721 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
      781 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
      841 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      901 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      961 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1021 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1081 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     1141 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1201 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1261 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1321 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1381 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1441 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1501 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1561 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1621 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1681 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1741 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1801 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1861 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1921 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1981 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2041 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2101 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2161 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2221 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2281 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2341 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2401 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2461 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2521 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2581 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2641 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2701 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2761 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2821 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2881 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2941 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3001 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3061 gcgcccaata c
//
