LOCUS       sequence-379023-        6488 bp    DNA     circular SYN 15-APR-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 6488)
  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 6488)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_BC16
FEATURES             Location/Qualifiers
     source          1..6488
                     /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"
     terminator      4699..4742
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4799..5387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4879..4898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5558..6349,6350..6418))
                     /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     6181..6200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6419..6482)
                     /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 cgtaggttaa gagctcaagc ttctgagatg caggtacatc ccactgatga
     4141 gtcccaaata ggacgaaacg cgcttcggtg cgtctgggat tccactgcta tccacgcggc
     4201 cgcacggcat tacgttagac gtctaggcca caccaccctg aaggcgcctg ctcgcctctg
     4261 atctgttgaa gctaagcagg gtcggtcctg gttagtactt ggatgggact ccgcctggta
     4321 atagccggta ccgtaggctt gcatggtcat agctgtttcc ttgcgctggg gatgcggtgg
     4381 gctctataga gagtcgatcg tgtgatcgaa ggaaccccta gtgatggagt tggccactcc
     4441 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc ggcctcagtg
     4501 agcgagcgag cgcgcagcat gtagagactg ggcctcatgg gccttccgcc tatcactgcc
     4561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa catggtcata gctgtttcct
     4621 tgcgtattgg gcgctctccg cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt
     4681 aaagcctggg gtgcctaaat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     4741 attgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5401 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5461 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5521 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5581 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5641 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5701 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5761 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5821 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5881 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5941 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6001 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6061 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6121 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6181 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6241 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6301 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6361 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6421 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6481 tatttgaa
//
