LOCUS Exported 7648 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 7648) 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 7648) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7648 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 858..885 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" CDS 2440..2457 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3242..3259) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3248..3472 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3518..3946 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3605..3624) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3815..3836 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3955..3975) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3960..4289 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4140..4275 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4202..4221 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4356..5150 /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(4410..4429) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5020..5039 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5324..5445 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5361..5380) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5415..5434 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5494..5510) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5494..5510) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5507..5529) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5518..5534 /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(5542..5572) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5587..5608 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5725..5742) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5896..6481) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5976..5995) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6652..7512) /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 7275..7294 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7513..7617) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gtccggattg acaattaatc atcggctcgt ataatatacc cgtttttttg 901 ggctagaaat aattttgtct aactttaaga aggagatatc tcgagatgtc ggtaccgctg 961 actacctcag cattcggcca cgcgtttctg gctaactgtg aacgtgagca gatccacctg 1021 gcgggctcca ttcagccgca cggtatcctg ctggctgtta aagagcctga caacgtggtg 1081 atccaggctt ctattaacgc tgcggagttc ctgaacacca actttgttgt tggccgtccg 1141 ctgcgtgacc tgggcggcga tctgcctttg cagatcctgc cgcacctgaa cggcccgctg 1201 cacctggctc cgatgaccct gcgttgtact gtgggttctc cgccgcgtcg tgtggactgt 1261 accattcatc gaccgtctaa cggcggcctg atcgtagaac tggaaccagc aaccaaggcc 1321 actaacattg cgccggctct ggtcggtgcg cttcatcgta tcacttcttc atcctccctg 1381 atgggcctgt gtgacgaaac cgcgactatt ttccgtgaga ttactggctt cgaccgtgtg 1441 atggtaatgc gtctcggcgc gcttgacgat ctgactgaag agcagatcgc agagattaaa 1501 gaggctttct ccctatttga caaggacggg gacgggacga taacaaccaa ggagctgggg 1561 acggtgttcc ggtctctggg gcagaacccc acagaagcag agctgcagga catgatcaat 1621 gaagtagatg ccgatggcga cggcacattc gacttccctg agttcctgac gatgatggca 1681 aggaaaatga atgactcaga cagtgaagag gaaattagag aagcgttccg cgtgtttgat 1741 aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac agaccttggt 1801 gagaagttaa ctgatgagga ggttgatgaa atgatcaggg tagcagacaa cgatggggat 1861 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagggtgg cggaggttct 1921 gtagattcat cacgtcgtaa gtggaataag gcaggtcacg cagtcagagc tataggtcgg 1981 ctgagctcgc gtaggcatga tttgctgtcc gaatgtcgtc gtgcggacct ggaagcgttc 2041 ctgggtaacc gctacccggc gtctactatt ccgcagatcg ctcgtcgcct gtacgaactt 2101 aaccgtgttc gcctgctggt agatgtgaac tatactccgg ttccgctaca gccgcgcatc 2161 agcccgctga acggtcgtga tctggatatg tccctgtctt gcctgcgctc tatgtccccg 2221 atccaccaga aatacatgca ggacatgggc gttggcgcaa ccctggtttg ctctctgatg 2281 gtgtctggtc gtctgtgggg tctgatcgtt tgccaccact acgaaccgcg cttcgttccg 2341 tcccacattc gcgctgctgg cgaagcgctg gcggaaactt gtgcgaaccg catcgcgacg 2401 ctggagagct ttgcacagtc tcagtccaaa tgaaagcttc atcatcacca tcaccattga 2461 aagaaggaga tatacatacc catgagtgtc aacttagctt cccagttgcg ggaagggacg 2521 aaaaaatccc actccatggc ggagaacgtc ggctttgtca aatgcttcct caagggcgtt 2581 gtcgagaaaa attcctaccg taagctggtt ggcaatctct actttgtcta cagtgccatg 2641 gaagaggaaa tggcaaaatt taaggaccat cccatcctca gccacattta cttccccgaa 2701 ctcaaccgca aacaaagcct agagcaagac ctgcaattct attacggctc caactggcgg 2761 caagaagtga aaatttctgc cgctggccaa gcctatgtgg accgagtccg gcaagtggcc 2821 gctacggccc ctgaattgtt ggtggcccat tcctacaccc gttacctggg ggatctttcc 2881 ggcggtcaaa ttctcaagaa aattgcccaa aatgccatga atctccacga tggtggcaca 2941 gctttctatg aatttgccga cattgatgac gaaaaggctt ttaaaaatac ctaccgtcaa 3001 gctatgaatg atctgcccat tgaccaagcc accgccgaac ggattgtgga tgaagccaat 3061 gacgcctttg ccatgaacat gaaaatgttc aacgaacttg aaggcaacct gatcaaggcg 3121 atcggcatta tggtgttcaa cagcctcacc cgtcgccgca gtcaaggcag caccgaagtt 3181 ggcctcgcca cctccgaagg ctagtctaga tctagagggc ccgtttaaac ccgctgatca 3241 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 3301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 3361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 3421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 3481 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 4201 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 4261 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg 4321 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 4381 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 4441 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 4501 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 4561 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 4621 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 4681 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 4741 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 4801 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 4861 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 4921 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 4981 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 5041 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 5101 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct 5161 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 5221 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 5281 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc 5341 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 5401 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc 5461 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 5521 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 5581 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 5641 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 5701 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 5761 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 5821 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 5881 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 5941 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 6001 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 6061 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 6121 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 6181 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 6241 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 6301 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 6361 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 6421 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6481 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 6541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 6601 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 6661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 6721 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 6781 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6841 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6901 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6961 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7021 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7081 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7141 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7201 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7261 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7321 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 7381 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7441 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7501 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 7561 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 7621 atttccccga aaagtgccac ctgacgtc //