LOCUS Exported 7643 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial and mammalian expression plasmid for near-infrared fluorescent Ca2+ indicator NIR-GECO1. ACCESSION . VERSION . KEYWORDS pDuEx2-NIR-GECO1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7643) AUTHORS Qian Y, Piatkevich KD, Mc Larney B, Abdelfattah AS, Mehta S, Murdock MH, Gottschalk S, Molina RS, Zhang W, Chen Y, Wu J, Drobizhev M, Hughes TE, Zhang J, Schreiter ER, Shoham S, Razansky D, Boyden ES, Campbell RE TITLE A genetically encoded near-infrared fluorescent calcium ion indicator. JOURNAL Nat Methods. 2019 Feb;16(2):171-174. doi: 10.1038/s41592-018-0294-6. Epub 2019 Jan 21. PUBMED 30664778 REFERENCE 2 (bases 1 to 7643) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7643 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 2204..2221 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3000..3017) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3006..3230 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3276..3704 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3363..3382) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3573..3594 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3713..3733) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3718..4047 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3898..4033 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3960..3979 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4114..4908 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4168..4187) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4778..4797 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5082..5203 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5119..5138) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5173..5192 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5252..5268) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5252..5268) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5265..5287) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5276..5292 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5300..5330) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5345..5366 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5483..5500) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5654..6242) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5734..5753) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6413..7273) /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 7036..7055 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7274..7378) /gene="bla" /label=AmpR promoter primer_bind complement(7453..7472) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggtcc ggattcaatt aatcatcggc tcgtataata tacccgtttt 661 tttgggctag aaataatttt gtctaacttt aagaaggaga tatctcgaga tgtcggtacc 721 gctgactacc tcagcattcg gccacgcgtt tctggctaac tgtgaacgtg agcagatcca 781 cctggcgggc tccattcagc cgcacggtat cctgctggct gttaaagagc ctgacaacgt 841 ggtgatccag gcttctatta acgctgcgga gttcctgaac accaactttg ttgttggccg 901 tccgctgcgt gacctgggcg gcgatctgcc tttgcagatc ctgccgcacc tgaacggccc 961 gctgcacctg gctccgatga ccctgcgttg tactgtgggt tctccgccgc gtcgtgtgga 1021 ctgtaccatt catcgaccgt ctaacggcgg cctgatcgta gaactggaac cagcaaccaa 1081 ggccactaac attgcgccgg ctctggtcgg tgcgcttcat cgtatcactt cttcatcctc 1141 cctgatgggc ctgtgtgacg aaaccgcgac tattttccgt gagattactg gcttcgaccg 1201 tgtgatggta atgcgtctcg gcgcgcttga cgatctgact gaagagcaga tcgcagagat 1261 taaagaggct ttctccctat ttgacaagga cggggacggg acgataacaa ccaaggagct 1321 ggggacggtg ttccggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat 1381 caatgaagta gatgccgatg gcgacggcac attcgacttc cctgagttcc tgacgatgat 1441 ggcaaggaaa atgaatgact cagacagtga agaggaaatt agagaagcgt tccgcgtgtt 1501 tgataaggac ggcaatggct acatcggcgc agcagagctt cgccacgtga tgacagacct 1561 tggtgagaag ttaactgatg aggaggttga tgaaatgatc agggtagcag acaacgatgg 1621 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagg gtggcggagg 1681 ttctgtagat tcatcacgtc gtaagtggaa taaggcaggt cacgcagtca gagctatagg 1741 tcggctgagc tcgcgtaggc atgatttgct gtccgaatgt cgtcgtgcgg acctggaagc 1801 gttcctgggt aaccgctacc cggcgtctac tattccgcag atcgctcgtc gcctgtacga 1861 acttaaccgt gttcgcctgc tggtagatgt gaactatact ccggttccgc tacagccgcg 1921 catcagcccg ctgaacggtc gtgatctgga tatgtccctg tcttgcctgc gctctatgtc 1981 cccgatccac cagaaataca tgcaggacat gggcgttggc gcaaccctgg tttgctctct 2041 gatggtgtct ggtcgtctgt ggggtctgat cgtttgccac cactacgaac cgcgcttcgt 2101 tccgtcccac attcgcgctg ctggcgaagc gctggcggaa acttgtgcga accgcatcgc 2161 gacgctggag agctttgcac agtctcagtc caaatgaaag cttcatcatc accatcacca 2221 ttgaaagaag gagatataca tacccatgag tgtcaactta gcttcccagt tgcgggaagg 2281 gacgaaaaaa tcccactcca tggcggagaa cgtcggcttt gtcaaatgct tcctcaaggg 2341 cgttgtcgag aaaaattcct accgtaagct ggttggcaat ctctactttg tctacagtgc 2401 catggaagag gaaatggcaa aatttaagga ccatcccatc ctcagccaca tttacttccc 2461 cgaactcaac cgcaaacaaa gcctagagca agacctgcaa ttctattacg gctccaactg 2521 gcggcaagaa gtgaaaattt ctgccgctgg ccaagcctat gtggaccgag tccggcaagt 2581 ggccgctacg gcccctgaat tgttggtggc ccattcctac acccgttacc tgggggatct 2641 ttccggcggt caaattctca agaaaattgc ccaaaatgcc atgaatctcc acgatggtgg 2701 cacagctttc tatgaatttg ccgacattga tgacgaaaag gcttttaaaa atacctaccg 2761 tcaagctatg aatgatctgc ccattgacca agccaccgcc gaacggattg tggatgaagc 2821 caatgacgcc tttgccatga acatgaaaat gttcaacgaa cttgaaggca acctgatcaa 2881 ggcgatcggc attatggtgt tcaacagcct cacccgtcgc cgcagtcaag gcagcaccga 2941 agttggcctc gccacctccg aaggctaatc tagagggccc gtttaaaccc gctgatcagc 3001 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3061 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3121 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3181 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3241 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3421 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3481 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 3541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 3601 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 3661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 3721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 3841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 3901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 3961 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4021 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4081 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4141 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4201 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4261 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4321 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4381 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 4441 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 4501 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4561 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 4621 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 4681 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4741 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4801 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 4861 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 4921 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4981 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5041 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5101 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5221 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5401 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5461 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5521 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 5881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6061 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6361 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 7021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 7081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 7141 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 7201 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7261 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7381 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 7441 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc 7501 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg 7561 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga 7621 tgtacgggcc agatatacgc gtt //