LOCUS       sequence-307160-        5026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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   Moudgil A, Wilkinson MN, Chen X, He J, Cammack AJ, Vasek MJ, Lagunas
            T Jr, Qi Z, Lalli MA, Guo C, Morris SA, Dougherty JD, Mitra RD
  TITLE     Self-Reporting Transposons Enable Simultaneous Readout of Gene 
            Expression and Transcription Factor Binding in Single Cells.
  JOURNAL   Cell. 2020 Aug 20;182(4):992-1008.e21. doi: 
            10.1016/j.cell.2020.06.037. Epub 2020 Jul 24.
  PUBMED    32710817
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"; doi: 
            "10.1016/j.cell.2020.06.037"; journalName: "Cell"; date: 
            "2020-08-20- 20"; volume: "182"; issue: "4"; pages: "992-1008.e21"
COMMENT     Sequence Label: PB-SRT-Puro
FEATURES             Location/Qualifiers
     source          1..5026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     700..720
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             766..1365
                     /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(766..785)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1262..1282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     repeat_region   1743..1777
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2061..2078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2171..2759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2251..2270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2930..3721,3722..3790))
                     /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     3553..3572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3791..3895)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4196..4218
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4210..4227
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4211..4227
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4238..4257
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4238..4256
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   4292..4354
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        join(4596..5026,1..752)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(4827..5026,1..743)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
       61 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      121 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      181 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
      241 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
      301 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
      361 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
      421 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
      481 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
      541 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
      601 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
      661 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
      721 aaagtttttt tcttccattt caggtgtcgt gaaaactacg ccaccatgac cgagtacaag
      781 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
      841 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
      901 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
      961 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     1021 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     1081 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     1141 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     1201 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     1261 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     1321 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgataaca actcgagtac
     1381 cggcttaagt gatagatcgg aagagcggtt cagcaggaat gccgagaccg gtttaaactc
     1441 agtgtccata gcgacgccgg cgcgataagc tatctataac aagaaaatat atatataata
     1501 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     1561 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     1621 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     1681 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     1741 tgcatgcgtc aattttacgc agactatctt tctagggtta agagctcctg agatgcaggt
     1801 acatcccact gatgagtccc aaataggacg aaacgcgctt cggtgcgtct gggattccac
     1861 tgctatccac gcggccgcac ggcattacgt tagacgtcta ggccacacca ccctgaaggc
     1921 gcctgctcgc ctctgatctg ttgaagctaa gcagggtcgg tcctggttag tacttggatg
     1981 ggactccgcc tggtaatagc cggtaccgta ggcttgcatg gtcatagctg tttccttgcg
     2041 tattgggcgc tctccgcttc ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag
     2101 cctggggtgc ctaatgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2161 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2221 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2281 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2341 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2401 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2461 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2521 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2581 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2641 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2701 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2761 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2821 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2881 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2941 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3001 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3061 tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3121 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3181 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3241 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3301 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3361 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3421 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3481 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3541 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3601 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3661 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3721 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3781 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3841 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3901 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3961 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4021 atcaaaagaa tagaccgaga tagggttgag tggccgctac agggcgctcc cattcgccat
     4081 tcaggctgcg caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag
     4141 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     4201 tcacgacgtt gtaaaacgac ggccagtgag cgcgacgtaa tacgactcac tatagggcga
     4261 attggcggaa ggccgtcaag gccgcattta accctagaaa gataatcata ttgtgacgta
     4321 cgttaaagat aatcatgtgt aaaattgacg catgtgtttt atcggtctgt atatcgaggt
     4381 ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca tactaataat
     4441 aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaacaa aaactcaaaa
     4501 tttcttctat aaagtaacaa aacttttatc gaattcctgc agcccagctt gatcttaggt
     4561 ccctcgacga attctctaga gaagagcatg cgtgaggctc cggtgcccgt cagtgggcag
     4621 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     4681 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     4741 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     4801 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     4861 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacgcccctg gctgcagtac
     4921 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     4981 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ttgggc
//
