LOCUS Exported 6447 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION ABA-inducible split T7 gRNA for GFP vector using RNAPN d5-19 with CGG RNAP-C. ACCESSION . VERSION . KEYWORDS pJin-290 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6447) AUTHORS Pu J, Kentala K, Dickinson BC TITLE Multidimensional Control of Cas9 by Evolved RNA Polymerase-Based Biosensors. JOURNAL ACS Chem Biol. 2018 Feb 16;13(2):431-437. doi: 10.1021/acschembio.7b00532. Epub 2017 Aug 22. PUBMED 28809467 REFERENCE 2 (bases 1 to 6447) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6447 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 465..748 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers" promoter 985..1084 /label=CMVd1 promoter /note="truncated human cytomegalovirus (CMV) immediate early promoter that yields slightly reduced expression relative to the full-length promoter" CDS 1128..1451 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL LKLE" promoter 3648..3747 /label=CMVd1 promoter /note="truncated human cytomegalovirus (CMV) immediate early promoter that yields slightly reduced expression relative to the full-length promoter" primer_bind complement(4667..4685) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 4722..4843 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4733..4752) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4787..4806 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS 5242..6036 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5296..5315) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5906..5925 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(6208..349) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6288..6307) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 61 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 181 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt 361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 421 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac 481 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga 541 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat 601 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct 661 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc 721 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatggaaa 781 aaaactaaca gatactctag tagaaaaaaa ctaacagata ctctagtaaa aaaaatctcg 841 ccaacaagtt gacgagataa acacggcatt ttgccttgtt ttagtagatt ctgtaatttt 901 cattacagag tactaaaact atagggagag ccctcgaact cctatagtga ccggtattac 961 gtccggcgta gaggatcgag atctccaaaa tcaacgggac tttccaaaat gtcgtaataa 1021 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 1081 agctggttta gtgaaccgtc agatcactag aagcttacgc taccaccatg ggagtgcagg 1141 tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg 1201 tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca 1261 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg 1321 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg 1381 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc 1441 taaaactgga aacgtcgggt ggaagcggta aagcatttat gcaagttgtc gaggctgaca 1501 tgctctctaa gggtctactc ggtggcgagg cgtggtcttc gtggcataag gaagactcta 1561 ttcatgtagg agtacgctgc atcgagatgc tcattgagtc aaccggaatg gttagcttac 1621 accgccaaaa tgctggcgta gtaggtcaag actctgagac tatcgaactc gcacctgaat 1681 acgctgaggc tatcgcaacc cgtgcaggtg cgctggctgg catctctccg atgttccaac 1741 cttgcgtagt tcctcctaag ccgtggactg gcattactgg tggtggctat tgggctaacg 1801 gtcgtcgtcc tctggcgctg gtgcgtactc acagtaagaa agcactgatg cgctacgaag 1861 acgtttacat gcctgaggtg tacaaagcga ttaacattgc gcaaaacacc gcatggaaaa 1921 tcaacaagaa agtcctagcg gtcgccaacg taatcaccaa gtggaagcat tgtccggtcg 1981 aggacatccc tgcgattgag cgtgaagaac tcccgatgaa accggaagac atcgacatga 2041 atcctgaggc tctcaccgcg tggaaacgtg ctgccgctgc tgtgtaccgc aaggacaagg 2101 ctcgcaagtc tcgccgtatc agccttgagt tcatgcttga gcaagccaat aagtttgcta 2161 accataaggc catctggttc ccttacaaca tggactggcg cggtcgtgtt tacgctgtgt 2221 caatgttcaa cccgcaaggt aacgatatga ccaaaggact gcttacgctg gcgaaaggta 2281 aaccaatcgg taaggaaggt tactactggc tgaaaatcca cggtgcaaac tgtgcgggtg 2341 tcgataaggt tccgttccct gagcgcatca agttcattga ggaaaaccac gagaacatca 2401 tggcttgcgc taagtctcca ctggagaaca cttggtgggc tgagcaagat tctccgttct 2461 gcttccttgc gttctgcttt gagtacgctg gggtacagca ccacggcctg agctataact 2521 gctcccttcc gctggcgttt gacgggtctt gctctggcat ccagcacttc tccgcgatgc 2581 tccgagatga ggtaggtggt cgcgcggtta acttgcttcc tagtgaaacc gttcaggaca 2641 tctacgggat tgttgctaag aaagtcaacg agattctaca agcagacgca atcaatggga 2701 ccgataacga agtagttacc gtgaccgatg agaacactgg tgaaatctct gagaaagtca 2761 agctgggcac taaggcactg gctggtcaat ggctggctta cggtgttact cgcagtgtga 2821 ctaagcgttc agtcatgacg ctggcttacg ggtccaaaga gttcggcttc cgtcaacaag 2881 tgctggaaga taccattcag ccagctattg attccggcaa gggtctgatg ttcactcagc 2941 cgaatcaggc tgctggatac atggctaagc tgatttggga atctgtgagc gtgacggtgg 3001 tagctgcggt tgaagcaatg aactggctta agtctgctgc taagctgctg gctgctgagg 3061 tcaaagataa gaagactgga gagattcttc gcaagcgttg cgctgtgcat tgggtaactc 3121 ctgatggttt ccctgtgtgg caggaataca agaagcctat taaaacgcgc gtgcatatta 3181 tgttcctcgg tcagttcgaa atgcagccta ccattaacac caacaaagat agcgagattg 3241 atgcacgcaa acaggagtct ggtatcgctc ctaactttgt acacagccaa gacggtagcc 3301 accttcgtaa gactgtagtg tgggcacacg agaagtacgg aatcgaatct tttgcactga 3361 ttcacgactc cttcggtacc attccggctg acgctgcgaa cctgttcaaa gcagtgcgcg 3421 aaactatggt tgacacatat gagtcttgtg atgtactggc tgatttctac gaccagttcg 3481 ctgaccagtt gcacgagtct caattggaca aaatgccagc acttccggct aaaggtaact 3541 tgaacctccg tgacatctta gagtcggact tcgcgttcgc gtaatgttta aacgaattcg 3601 ggctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagcca aaatcaacgg 3661 gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta 3721 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcac tagaagctta 3781 cgctaccacc atgaacacga ttaacatcgc taagaacgac ttctctgaca tcgaactggc 3841 tgctatcccg ttaaacactc tggctgacca ttacggtgag cgttcagctc gcggacagtt 3901 ggcccttgag catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagtg 3961 tcaacttaaa gctggtaagg ttgcggataa tgctgccgcc aagcctctca tcactaccct 4021 actccctaag atgattgccc gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg 4081 caggcgcccg acagccttca agttcctaaa agaaatcaag ccggaagccg tagcgtacat 4141 caccattaag acctctctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt 4201 agcaagcgca atcggtcgga ccattgagga cgaggctcgc ttcggtcgta tccgtgacct 4261 tgaagctaag cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt 4321 ctacaagggt ggctctggct ctggctcgag cgcggtgctt tggcatgagc aatggtatga 4381 gggtctggat gacgccagta ggcagttttt tggagaacat aataccgaaa aaatgtttgc 4441 tgctttagag cctctgtacg aaatgctgaa gagaggaccg gaaactttga gggaaatatc 4501 gttccaaaat tcttttggta gggacttgaa tgacgcttac gaatggctga tgaattacaa 4561 aaaatctaaa gatgttagta atttaaacca agcgtgggac atttactata atgttttcag 4621 gaaaattggt taaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa 4681 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt 4741 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 4801 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 4861 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct 4921 ctacaaatgt ggtaaaatcg aattctaatg gatcctcttt gcgcttgcgt tttcccttgt 4981 ccagatagcc cagtagctga cattcatccg gggtcagcac catttctgcg gactggcttt 5041 ctacgtgttc cgcttccttt agcagccctt gcgccctgag tgcttgcggc agcgtgagct 5101 tcaaaagaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac 5161 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag 5221 acaggatgac ggtcgtttcg catgcttgaa caagatggat tgcacgcagg ttctccggcc 5281 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 5341 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 5401 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 5461 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 5521 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 5581 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 5641 gaccaccaag cgaaacatcg catcgagcga gcacgcactc ggatggaagc cggtcttgtc 5701 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 5761 ctcaaggcgc gtatgccgga tggtgaggat ctcgtcgtga ctcatggcga tgcctgcttg 5821 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 5881 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 5941 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 6001 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 6061 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac 6121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6421 tctcagttcg gtgtaggtcg ttcgctc //