LOCUS       sequence-308644-        5859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac self-reporting transposon with tdTomato marker for 
            mammalian calling cards.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5859)
  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 5859)
  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-tdTomato
FEATURES             Location/Qualifiers
     source          1..5859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..125
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     117..134
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     118..134
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     145..164
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        145..163
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   199..261
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        503..1685
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          734..1676
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1633..1653
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1697..3127
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQAA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     repeat_region   3509..3543
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3827..3844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3937..4525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4017..4036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4696..5487,5488..5556))
                     /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     5319..5338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5557..5661)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgttgggaa gggcgtttcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
       61 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      121 aaacgacggc cagtgagcgc gacgtaatac gactcactat agggcgaatt ggcggaaggc
      181 cgtcaaggcc gcatttaacc ctagaaagat aatcatattg tgacgtacgt taaagataat
      241 catgtgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta tttattaatt
      301 tgaatagata ttaagtttta ttatatttac acttacatac taataataaa ttcaacaaac
      361 aatttattta tgtttattta tttattaaaa aaaacaaaaa ctcaaaattt cttctataaa
      421 gtaacaaaac ttttatcgaa ttcctgcagc ccagcttgat cttaggtccc tcgacgaatt
      481 ctctagagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      541 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      601 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      661 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      721 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      781 tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc
      841 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      901 cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg cgaatctggt
      961 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     1021 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     1081 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     1141 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     1201 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     1261 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     1321 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     1381 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1441 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1501 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1561 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1621 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1681 cgtgaaaact acgccaatgg tgagcaaggg cgaggaggtc atcaaagagt tcatgcgctt
     1741 caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     1801 gggccgcccc tacgagggca cccaggccgc caagctgaag gtgaccaagg gcggccccct
     1861 gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa
     1921 gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga
     1981 gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca
     2041 ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc
     2101 cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga
     2161 cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccactacct
     2221 ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg gctactacta
     2281 cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2341 cgagcgctcc gagggccgcc accacctgtt cctggggcat ggcaccggca gcaccggcag
     2401 cggcagctcc ggcaccgcct cctccgagga caacaacatg gccgtcatca aagagttcat
     2461 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     2521 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     2581 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     2641 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     2701 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     2761 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     2821 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     2881 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     2941 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     3001 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     3061 acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta
     3121 caagtaatcg ataacaactc gagtaccggc ttaagtgata gatcggaaga gcggttcagc
     3181 aggaatgccg agaccggttt aaactcagtg tccatagcga cgccggcgcg ataagctatc
     3241 tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata
     3301 taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga
     3361 ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca
     3421 cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt
     3481 agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta
     3541 gggttaagag ctcctgagat gcaggtacat cccactgatg agtcccaaat aggacgaaac
     3601 gcgcttcggt gcgtctggga ttccactgct atccacgcgg ccgcacggca ttacgttaga
     3661 cgtctaggcc acaccaccct gaaggcgcct gctcgcctct gatctgttga agctaagcag
     3721 ggtcggtcct ggttagtact tggatgggac tccgcctggt aatagccggt accgtaggct
     3781 tgcatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact
     3841 cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa
     3901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4801 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     4861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     5701 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5761 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtggc
     5821 cgctacaggg cgctcccatt cgccattcag gctgcgcaa
//
