LOCUS Exported 7276 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION ABA-inducible split T7 gRNA for GFP vector using RNAPN d5-19. ACCESSION . VERSION . KEYWORDS pJin-264 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7276) 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 7276) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7276 /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" primer_bind complement(944..963) /label=T7 /note="T7 promoter, forward primer" promoter complement(945..963) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 989..1088 /label=CMVd1 promoter /note="truncated human cytomegalovirus (CMV) immediate early promoter that yields slightly reduced expression relative to the full-length promoter" promoter 3862..3961 /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(5496..5514) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 5551..5672 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5562..5581) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5616..5635 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS 6071..6865 /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(6125..6144) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6735..6754 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(7037..349) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7117..7136) /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 tactaaaacg ccgaggtgaa gttcgagggc tctccctata gtgagtcgta 961 ttacgtccgg cgtagaggat cgagatctcc aaaatcaacg ggactttcca aaatgtcgta 1021 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 1081 gcatagctgg tttagtgaac cgtcagatca ctagaagctt acgctaccac catgacccag 1141 gacgagttta cccagctgtc acagtcaatc gccgagtttc acacctatca gctggggaac 1201 ggaagatgta gtagtctgct ggcccagaga atccacgctc cccctgaaac cgtgtggagc 1261 gtggtccgga gattcgatag accacaaatc tacaagcact tcatcaagag ctgcaacgtg 1321 tccgaagatt tcgagatgcg cgtcggatgt acacgagacg tgaacgtcat ctctggcctg 1381 cccgcaaata ctagtcgaga gcggctggac ctgctggacg atgacaggcg cgtgactggg 1441 ttctccatta ccggcgggga acacagactg aggaactaca agtctgtgac cacagtccat 1501 aggtttgaga aagaggaaga ggaagagcgc atttggaccg tggtcctgga aagctacgtg 1561 gtcgatgtgc ccgagggcaa tagcgaagag gacacaaggc tgtttgccga tactgtgatt 1621 cggctgaacc tgcagaaact ggcaagcatt actgaggcta tgaacacgtc gggtggaagc 1681 ggtaaagcat ttatgcaagt tgtcgaggct gacatgctct ctaagggtct actcggtggc 1741 gaggcgtggt cttcgtggca taaggaagac tctattcatg taggagtacg ctgcatcgag 1801 atgctcattg agtcaaccgg aatggttagc ttacaccgcc aaaatgctgg cgtagtaggt 1861 caagactctg agactatcga actcgcacct gaatacgctg aggctatcgc aacccgtgca 1921 ggtgcgctgg ctggcatctc tccgatgttc caaccttgcg tagttcctcc taagccgtgg 1981 actggcatta ctggtggtgg ctattgggct aacggtcgtc gtcctctggc gctggtgcgt 2041 actcacagta agaaagcact gatgcgctac gaagacgttt acatgcctga ggtgtacaaa 2101 gcgattaaca ttgcgcaaaa caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc 2161 aacgtaatca ccaagtggaa gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa 2221 gaactcccga tgaaaccgga agacatcgac atgaatcctg aggctctcac cgcgtggaaa 2281 cgtgctgccg ctgctgtgta ccgcaaggac aaggctcgca agtctcgccg tatcagcctt 2341 gagttcatgc ttgagcaagc caataagttt gctaaccata aggccatctg gttcccttac 2401 aacatggact ggcgcggtcg tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat 2461 atgaccaaag gactgcttac gctggcgaaa ggtaaaccaa tcggtaagga aggttactac 2521 tggctgaaaa tccacggtgc aaactgtgcg ggtgtcgata aggttccgtt ccctgagcgc 2581 atcaagttca ttgaggaaaa ccacgagaac atcatggctt gcgctaagtc tccactggag 2641 aacacttggt gggctgagca agattctccg ttctgcttcc ttgcgttctg ctttgagtac 2701 gctggggtac agcaccacgg cctgagctat aactgctccc ttccgctggc gtttgacggg 2761 tcttgctctg gcatccagca cttctccgcg atgctccgag atgaggtagg tggtcgcgcg 2821 gttaacttgc ttcctagtga aaccgttcag gacatctacg ggattgttgc taagaaagtc 2881 aacgagattc tacaagcaga cgcaatcaat gggaccgata acgaagtagt taccgtgacc 2941 gatgagaaca ctggtgaaat ctctgagaaa gtcaagctgg gcactaaggc actggctggt 3001 caatggctgg cttacggtgt tactcgcagt gtgactaagc gttcagtcat gacgctggct 3061 tacgggtcca aagagttcgg cttccgtcaa caagtgctgg aagataccat tcagccagct 3121 attgattccg gcaagggtct gatgttcact cagccgaatc aggctgctgg atacatggct 3181 aagctgattt gggaatctgt gagcgtgacg gtggtagctg cggttgaagc aatgaactgg 3241 cttaagtctg ctgctaagct gctggctgct gaggtcaaag ataagaagac tggagagatt 3301 cttcgcaagc gttgcgctgt gcattgggta actcctgatg gtttccctgt gtggcaggaa 3361 tacaagaagc ctattcagac gcgcttgaac ctgatgttcc tcggtcagtt ccgcttacag 3421 cctaccatta acaccaacaa agatagcgag attgatgcac acaaacagga gtctggtatc 3481 gctcctaact ttgtacacag ccaagacggt agccaccttc gtaagactgt agtgtgggca 3541 cacgagaagt acggaatcga atcttttgca ctgattcacg actccttcgg taccattccg 3601 gctgacgctg cgaacctgtt caaagcagtg cgcgaaacta tggttgacac atatgagtct 3661 tgtgatgtac tggctgattt ctacgaccag ttcgctgacc agttgcacga gtctcaattg 3721 gacaaaatgc cagcacttcc ggctaaaggt aacttgaacc tccgtgacat cttagagtcg 3781 gacttcgcgt tcgcgtaatg tttaaacgaa ttcgggctcg gtacccgggg atcctctaga 3841 gtcgacctgc aggcatgcaa gccaaaatca acgggacttt ccaaaatgtc gtaataaccc 3901 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 3961 tggtttagtg aaccgtcaga tcactagaag cttacgctac caccatgaac acgattaaca 4021 tcgctaagaa cgacttctct gacatcgaac tggctgctat cccgttaaac actctggctg 4081 accattacgg tgagcgttca gctcgcggac agttggccct tgagcatgag tcttacgaga 4141 tgggtgaagc acgcttccgc aagatgtttg agtgtcaact taaagctggt aaggttgcgg 4201 ataatgctgc cgccaagcct ctcatcacta ccctactccc taagatgatt gcccgcatca 4261 acgactggtt tgaggaagtg aaagctaagc gcggcaggcg cccgacagcc ttcaagttcc 4321 taaaagaaat caagccggaa gccgtagcgt acatcaccat taagacctct ctggcttgcc 4381 taaccagtgc tgacaataca accgttcagg ctgtagcaag cgcaatcggt cggaccattg 4441 aggacgaggc tcgcttcggt cgtatccgtg accttgaagc taagcacttc aagaaaaacg 4501 ttgaggaaca actcaacaag cgcgtagggc acgtctacaa gggtggctct ggctctggct 4561 cgagcgtccc cctgtatggg ttcaccagca tttgtgggcg gagaccagag atggaggcag 4621 ccgtcagcac tattccccgg ttcctgcagt caagctccgg cagcatgctg gacgggaggt 4681 tcgatccaca gtccgccgct cacttctttg gggtctacga tggacatggc gggtcccagg 4741 tggccaacta ttgcagggag cgcatgcacc tggctctggc agaggaaatc gccaaggaga 4801 aacctatgct gtgcgacgga gatacatggc tggaaaagtg gaagaaagcc ctgttcaact 4861 cttttctgcg ggtggactcc gagatcgaat ctgtcgctcc tgagaccgtg ggctctacaa 4921 gtgtggtcgc agtggtcttc ccatctcaca tttttgtcgc aaattgcggc gacagccggg 4981 ccgtgctgtg caggggaaag accgccctgc cactgagcgt ggaccataaa cccgacaggg 5041 aggatgaagc agcccgcatc gaggctgcag gaggcaaagt gatccagtgg aacggagcac 5101 gggtgtttgg cgtcctggcc atgtcacgca gcattgggga tcgatacctg aaaccatcaa 5161 tcattcccga ccctgaagtg actgccgtca agagagtgaa agaagacgat tgcctgatcc 5221 tggctagcga cggcgtctgg gatgtgatga ccgacgagga agcatgtgag atggcccgaa 5281 agcggattct gctgtggcat aagaaaaatg ccgtggctgg cgatgctagt ctgctggcag 5341 acgagcggag aaaggaaggg aaagatcccg ccgctatgag tgcagccgaa tatctgtcaa 5401 aactggctat tcagaggggc agtaaggaca acatctccgt cgtcgtcgtg gacctgaagt 5461 gaaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc 5521 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca 5581 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat 5641 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt 5701 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg 5761 gtaaaatcga attctaatgg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc 5821 agtagctgac attcatccgg ggtcagcacc atttctgcgg actggctttc tacgtgttcc 5881 gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgagctt caaaagaatt 5941 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt 6001 cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgacg 6061 gtcgtttcgc atgcttgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 6121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 6181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 6241 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 6301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 6361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 6421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 6481 gaaacatcgc atcgagcgag cacgcactcg gatggaagcc ggtcttgtcg atcaggatga 6541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg 6601 tatgccggat ggtgaggatc tcgtcgtgac tcatggcgat gcctgcttgc cgaatatcat 6661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 6721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 6781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 6841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg 6901 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 6961 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 7021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 7081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 7141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 7201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 7261 tgtaggtcgt tcgctc //