LOCUS       sequence-378969-        5026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded piggyBac self-reporting transposon with puromycin marker 
            for mammalian calling cards.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5026)
  AUTHORS   Lalli M, Yen A, Thopte U, Dong F, Moudgil A, Chen X, Milbrandt J, 
            Dougherty JD, Mitra RD
  TITLE     Measuring transcription factor binding and gene expression using 
            barcoded self-reporting transposon calling cards and transcriptomes.
  JOURNAL   NAR Genom Bioinform. 2022 Aug 31;4(3):lqac061. doi: 
            10.1093/nargab/lqac061. eCollection 2022 Sep.
  PUBMED    36062164
REFERENCE   2  (bases 1 to 5026)
  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: "NAR Genom 
            Bioinform."; date: "2022-08-31"; volume: "4(3):lqac061. doi"; pages:
            " 10.1093/nargab/lqac061. eCollection 2022 Sep"
COMMENT     Sequence Label: PB-SRT-Puro_BC23
FEATURES             Location/Qualifiers
     source          1..5026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   374..436
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        678..1860
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          909..1851
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1808..1828
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1874..2473
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1874..1893)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2370..2390
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     repeat_region   2851..2885
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3169..3186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3279..3867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3359..3378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4038..4829,4830..4898))
                     /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     4661..4680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4899..5003)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccgcatt taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgt
      421 gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat
      481 agatattaag ttttattata tttacactta catactaata ataaattcaa caaacaattt
      541 atttatgttt atttatttat taaaaaaaac aaaaactcaa aatttcttct ataaagtaac
      601 aaaactttta tcgaattcct gcagcccagc ttgatcttag gtccctcgac gaattctcta
      661 gagaagagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      721 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      781 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      841 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      901 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      961 cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct tgatcccgag
     1021 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
     1081 cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat ctggtggcac
     1141 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
     1201 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
     1261 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
     1321 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     1381 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     1441 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     1501 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     1561 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     1621 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     1681 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg
     1741 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     1801 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga
     1861 aaactacgcc accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     1921 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     1981 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     2041 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     2101 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     2161 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     2221 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     2281 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     2341 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     2401 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     2461 gcccggtgcc tgataacaac tcgagtaccg gcttaagtga tagatcggaa gagcggttca
     2521 gcaggaatgc cgagaccggt ttaaactcag tgtccatagc gacgccggcg cgataagcta
     2581 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa
     2641 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt
     2701 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct
     2761 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat
     2821 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc
     2881 tagggttaag agctcctgag atgcaggtac atcccactga tgagtcccaa ataggacgaa
     2941 acgcgcttcg gtgcgtctgg gattccactg ctatccacgc ggccgcacgg cattacgtta
     3001 gacgtctagg ccacaccacc ctgaaggcgc ctgctcgcct ctgatctgtt gaagctaagc
     3061 agggtcggtc ctggttagta cttggatggg actccgcctg gtaatagccg gtaccgtagg
     3121 cttgcatggt catagctgtt tccttgcgta ttgggcgctc tccgcttcct cgctcactga
     3181 ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa
     3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc accggctcca
     4201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
