LOCUS Exported 5026 bp ds-DNA circular SYN 06-DEC-2022 DEFINITION Barcoded piggyBac self-reporting transposon with puromycin marker for mammalian calling cards. ACCESSION . VERSION . KEYWORDS PB-SRT-Puro_BC23 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5026) 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 5026) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 6, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5026 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 278..300 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 292..309 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 293..309 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 320..339 /label=T7 /note="T7 promoter, forward primer" promoter 320..338 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" repeat_region 374..436 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" promoter 679..1860 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 909..1851 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1808..1828 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1874..2473 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1874..1893) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2370..2390 /label=Puro-F /note="Puromycin resistance gene, forward primer" repeat_region 2851..2885 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(3169..3186) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3279..3867) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3359..3378) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4038..4898) /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 4661..4680 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4899..5003) /gene="bla" /label=AmpR promoter ORIGIN 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt 181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca 361 aggccgcatt taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgt 421 gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat 481 agatattaag ttttattata tttacactta catactaata ataaattcaa caaacaattt 541 atttatgttt atttatttat taaaaaaaac aaaaactcaa aatttcttct ataaagtaac 601 aaaactttta tcgaattcct gcagcccagc ttgatcttag gtccctcgac gaattctcta 661 gagaagagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag 721 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg 781 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag 841 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca 901 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc 961 cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct tgatcccgag 1021 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct 1081 cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat ctggtggcac 1141 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct 1201 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact 1261 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt 1321 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct 1381 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca 1441 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct 1501 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca 1561 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg 1621 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg 1681 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg 1741 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg 1801 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga 1861 aaactacgcc accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 1921 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 1981 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 2041 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 2101 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 2161 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 2221 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 2281 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 2341 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 2401 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 2461 gcccggtgcc tgataacaac tcgagtaccg gcttaagtga tagatcggaa gagcggttca 2521 gcaggaatgc cgagaccggt ttaaactcag tgtccatagc gacgccggcg cgataagcta 2581 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa 2641 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt 2701 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct 2761 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat 2821 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc 2881 tagggttaag agctcctgag atgcaggtac atcccactga tgagtcccaa ataggacgaa 2941 acgcgcttcg gtgcgtctgg gattccactg ctatccacgc ggccgcacgg cattacgtta 3001 gacgtctagg ccacaccacc ctgaaggcgc ctgctcgcct ctgatctgtt gaagctaagc 3061 agggtcggtc ctggttagta cttggatggg actccgcctg gtaatagccg gtaccgtagg 3121 cttgcatggt catagctgtt tccttgcgta ttgggcgctc tccgcttcct cgctcactga 3181 ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa 3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 3841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 3901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 3961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 4021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 4081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 4141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc accggctcca 4201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 4261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 4321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 4381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 4441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 4501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 4561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 4621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 4681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 4741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 4801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 4861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 4921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 4981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac //