LOCUS Exported 5178 bp ds-DNA circular SYN 17-NOV-2021 DEFINITION Mammalian expression of green fluorescent potassium indicator KRaION2. ACCESSION . VERSION . KEYWORDS pKRaION2-N1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5178) AUTHORS Torres Caban CC, Yang M, Lai CC, Yang L, Subach F, Smith BO, Piatkevich KD, Boyden ES TITLE Tuning the sensitivity of genetically encoded fluorescent potassium indicators through structure-guided and genome mining strategies. JOURNAL bioRxiv 2021.10.07.463355 REFERENCE 2 (bases 1 to 5178) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5178 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 591..671 /label=MCS /note="multiple cloning site" polyA_signal 1966..2087 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2003..2022) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2057..2076 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2094..2549) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2231..2252) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2443..2462 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2576..2680 /gene="bla" /label=AmpR promoter promoter 2682..3039 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2705..2725) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2890..3025 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2952..2971 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3074..3868 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3128..3147) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3738..3757 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4056..4075) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4100..4147 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4476..5064 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4965..4984 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg 661 gatccaccgg tatggtctca aagggagagg aggaaaacat ggctagtctg cctgctaccc 721 acgaactgca tatttttggc tccataaatg gcatcgattt cgacatggta ggccaaggca 781 ccggtaatcc aaacgacggc tatgaggagc tgaacctgaa gtccaccatg ggcgatctgc 841 agttcagtcc ctggatcctg gtgccgcaca ttgggtacgg gtttcaccag tatcttccct 901 atccagacgg gatgtcacca tttcaggctg caatggtcga cgggtccgga taccaagtgc 961 accgcactat gcagtttgag gacggtgcct cactgaccgt gaattacaga tacacctacg 1021 aaggatcaca tatcaaaggg gaagctcagg tggagggcac tgggttccca gctgacggcc 1081 ctgtgatgac aaattccctg acagccgagg cccacggcct gttcaacttc gtgaaggacg 1141 ccggcgccaa gctgtgggat gccgtgacag gccagcacga caaggacgac caggccaaga 1201 aggtgcagga acacctgaac aagaccggca tccccgacgc cgacaaagtg aacatccaga 1261 tcgccgacgg caaggccaca gtgacaggcg acggactgag ccaggaagcc aaagaaaaga 1321 tcctggtggc cgtgggcaac atcagcggaa tcgccagcgt ggacgatcaa gtgaaaaccg 1381 ccacccctgc caccgccagc cagttctaca cagtgaagtc cggcgacacc ctgagcgcca 1441 tcagcaaaca ggtgtacggc aacgccaacc tgtacaacaa gattttcgag gccaacaagc 1501 ccatgctgaa gtcccccgac aaaatctacc ccggccaggt gctgagaatc cccgaggaaa 1561 tgtacttcgc tgactggtgc gtatccaaga agacttgccc caatgataag actattgtct 1621 ccacatttaa gtgggctttt atcaccgaca acggcaaacg ctaccggtct accgctcgca 1681 ccacatacac attcgccaag ccaatggccg caaattacct caaaaatcag ccgatgtatg 1741 tctttcggaa aacagagctg aaacattcta agaccgaact gaattttaaa gagtggcaaa 1801 aggcctttac ggacgtgatg ggcatggacg agctgtataa gtaagcggcc gcgactctag 1861 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 1921 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 1981 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 2041 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt 2101 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa 2161 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt 2221 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa 2281 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag 2341 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt 2401 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg 2461 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc 2521 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg 2581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 2641 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag 2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 2941 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg 3061 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 3121 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 3181 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 3241 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 3301 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 3361 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 3421 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 3481 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 3541 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 3601 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 3661 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 3721 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 3781 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 3841 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 3901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 3961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4021 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg 4081 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg 4141 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 4201 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 4261 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt 4321 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 4381 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 4441 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 4501 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 4561 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 4621 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 4681 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 4741 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 4801 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 4861 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 4921 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 4981 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 5041 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 5101 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 5161 cgtattaccg ccatgcat //