LOCUS       sequence-379116-        6504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded piggyBac self-reporting transposon with tdTomato marker in 
            AAV backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  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 6504)
  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: AAV-PB-SRT-tdTomato_BC23
FEATURES             Location/Qualifiers
     source          1..6504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..401
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     393..410
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     394..410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     421..440
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        421..439
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   476..605
                     /label=AAV2 ITR
     repeat_region   752..814
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1056..2238
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1287..2229
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2186..2206
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2250..3680
                     /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   4062..4096
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   4404..4533
                     /label=AAV2 ITR
     terminator      4715..4758
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4815..5403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4895..4914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5574..6365,6366..6434))
                     /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     6197..6216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6435..6498)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctaataaca ggcctgctgg taatcgcagg cctttttatt tggtgtattt agaaaaataa
       61 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
      121 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      181 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc
      241 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc
      301 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      361 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg
      421 taatacgact cactataggg cgaattgtag gcggaaggcc gtcaaggccg catggctgcg
      481 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      541 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      601 ttcctattga cgtcagatcg ctaaataatg acgtatgttc ccatagtaac gccaataggg
      661 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      721 caagtgtatc atacgtcaag gccgcattta accctagaaa gataatcata ttgtgacgta
      781 cgttaaagat aatcatgtgt aaaattgacg catgtgtttt atcggtctgt atatcgaggt
      841 ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca tactaataat
      901 aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaacaa aaactcaaaa
      961 tttcttctat aaagtaacaa aacttttatc gaattcctgc agcccagctt gatcttaggt
     1021 ccctcgacga attctctaga gaagagcatg cgtgaggctc cggtgcccgt cagtgggcag
     1081 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     1141 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     1201 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     1261 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     1321 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacgcccctg gctgcagtac
     1381 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     1441 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ttgggcgctg gggccgccgc
     1501 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     1561 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     1621 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     1681 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     1741 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     1801 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     1861 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     1921 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     1981 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     2041 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     2101 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     2161 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     2221 ccatttcagg tgtcgtgaaa actacgccaa tggtgagcaa gggcgaggag gtcatcaaag
     2281 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     2341 agggcgaggg cgagggccgc ccctacgagg gcacccaggc cgccaagctg aaggtgacca
     2401 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     2461 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     2521 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     2581 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     2641 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     2701 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     2761 gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     2821 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     2881 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg catggcaccg
     2941 gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac atggccgtca
     3001 tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg
     3061 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     3121 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg
     3181 gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc
     3241 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga
     3301 cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca
     3361 acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg
     3421 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga
     3481 aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc
     3541 aactgcccgg ctactactac gtggacacca agctggacat cacctcccac aacgaggact
     3601 acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc ctgtacggca
     3661 tggacgagct gtacaagtaa tcgataacaa ctcgagtacc ggcttaagtg atagatcgga
     3721 agagcggttc agcaggaatg ccgagaccgg tttaaactca gtgtccatag cgacgccggc
     3781 gcgataagct atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca
     3841 tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca
     3901 tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga
     3961 caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg
     4021 attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca
     4081 gactatcttt ctagggttaa gagctcctga gatgcaggta catcccactg atgagtccca
     4141 aataggacga aacgcgcttc ggtgcgtctg ggattccact gctatccacg cggccgcacg
     4201 gcattacgtt agacgtctag gccacaccac cctgaaggcg cctgctcgcc tctgatctgt
     4261 tgaagctaag cagggtcggt cctggttagt acttggatgg gactccgcct ggtaatagcc
     4321 ggtaccgtag gcttgcatgg tcatagctgt ttccttgcgc tggggatgcg gtgggctcta
     4381 tagagagtcg atcgtgtgat cgaaggaacc cctagtgatg gagttggcca ctccctctct
     4441 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4501 ccgggcggcc tcagtgagcg agcgagcgcg cagcatgtag agactgggcc tcatgggcct
     4561 tccgcctatc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaacatg
     4621 gtcatagctg tttccttgcg tattgggcgc tctccgcttc ctcgctcact gactcgctgc
     4681 gctcggtcgt tcgggtaaag cctggggtgc ctaaataaaa cgaaaggccc agtcttccga
     4741 ctgagccttt cgttttattg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5641 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5701 gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5761 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5821 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5881 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5941 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6001 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6061 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6181 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6481 ctcatgagcg gatacatatt tgaa
//
