LOCUS Exported 6506 bp ds-DNA circular SYN 07-DEC-2022 DEFINITION Barcoded piggyBac self-reporting transposon with tdTomato marker in AAV backbone. ACCESSION . VERSION . KEYWORDS AAV-PB-SRT-tdTomato_BC20 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6506) 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 6506) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 7, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6506 /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 4406..4535 /label=AAV2 ITR primer_bind complement(4663..4680) /label=L4440 /note="L4440 vector, forward primer" terminator 4717..4760 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(4817..5405) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4897..4916) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5576..6436) /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 6199..6218 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6437..6500) /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 tcagggttaa gagctcatct gagatgcagg tacatcccac tgatgagtcc 4141 caaataggac gaaacgcgct tcggtgcgtc tgggattcca ctgctatcca cgcggccgca 4201 cggcattacg ttagacgtct aggccacacc accctgaagg cgcctgctcg cctctgatct 4261 gttgaagcta agcagggtcg gtcctggtta gtacttggat gggactccgc ctggtaatag 4321 ccggtaccgt aggcttgcat ggtcatagct gtttccttgc gctggggatg cggtgggctc 4381 tatagagagt cgatcgtgtg atcgaaggaa cccctagtga tggagttggc cactccctct 4441 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 4501 gcccgggcgg cctcagtgag cgagcgagcg cgcagcatgt agagactggg cctcatgggc 4561 cttccgccta tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca 4621 tggtcatagc tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct 4681 gcgctcggtc gttcgggtaa agcctggggt gcctaaataa aacgaaaggc ccagtcttcc 4741 gactgagcct ttcgttttat tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 4801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 4861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 4921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 4981 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 5041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 5101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 5161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 5221 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 5401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 5461 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 5521 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 5581 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5641 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 5701 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc 5761 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 5821 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 5881 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 5941 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 6001 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 6061 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 6121 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 6181 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 6241 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 6301 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 6361 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6421 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 6481 gtctcatgag cggatacata tttgaa //