LOCUS Exported 6510 bp ds-DNA circular SYN 06-DEC-2022 DEFINITION Barcoded piggyBac self-reporting transposon with tdTomato marker in AAV backbone. ACCESSION . VERSION . KEYWORDS AAV-PB-SRT-tdTomato_BC10 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6510) 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 6510) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 6, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6510 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1057..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 4410..4539 /label=AAV2 ITR primer_bind complement(4667..4684) /label=L4440 /note="L4440 vector, forward primer" terminator 4721..4764 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(4821..5409) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4901..4920) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5580..6440) /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 6203..6222 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6441..6504) /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 catgggttaa 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 gacgcccggg 4501 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc atgtagagac tgggcctcat 4561 gggccttccg cctatcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 4621 aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact 4681 cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa ataaaacgaa aggcccagtc 4741 ttccgactga gcctttcgtt ttattgagca aaaggccagc aaaaggccag gaaccgtaaa 4801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 4861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 4921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 4981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 5041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 5101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 5161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 5221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 5281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 5341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 5401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 5461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 5521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 5581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 5641 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 5701 agtgctgcaa tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac 5761 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 5821 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 5881 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 5941 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 6001 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 6061 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 6121 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 6181 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 6241 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 6301 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 6361 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 6421 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 6481 tattgtctca tgagcggata catatttgaa //