LOCUS Exported 8662 bp ds-DNA circular SYN 22-MAY-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS CMV_OMM_jRCaMP1b_P2A-Mito4xGCaMP6f SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8662) AUTHORS . TITLE Direct Submission JOURNAL Exported May 22, 2019 from SnapGene 4.2.11 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8662 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 308..818 /label=CMV_Promoter /note="color: #ffffff; direction: RIGHT" primer_bind 769..789 /label=CMV_fwd_primer /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG" promoter 863..881 /label=T7_promoter /note="color: #ffffff" CDS 907..1005 /codon_start=1 /label=N-ter TOM20 33aa /note="color: #993366" /translation="MVGRNSAIAAGVCGALFIGYCIYFDRKRRSDPN" CDS 1006..1014 /codon_start=1 /label=linker /note="color: #c0c0c0" /translation="TGS" CDS 1015..2331 /codon_start=1 /product="no Nuclear Export Sequence" /label=jRCaMP1b /note="color: #ff0000" /translation="HHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKWGH AVRAIGRLSSANNTEMMYPADGGLRGYTHMALKVDGGGHLSCSFVTTYRSKKTVGNIKM PAIHYVSHRLERLEESDNEMFVVQREHAVAKFVGLGGGGGTGGSMNSLIKENMRMKVVL EGSVNGHQFKCTGEGEGNPYMGTQTMRIKVIEGGPLPFAFDILATSFMYGSRTFIKYPK GIPDFFKQSFPEGFTWERVTRYEDGGVITVMQDTSLEDGCLVYHVQVRGVNFPSNGAVM QKKTKGWEPTDSQLTEEQIAEFKEAFSLFDKDGDGTITTKEMGTVMRSLGQNPTEAELQ DMINEVDADGDGTIDFPEFLIMMAGKMKYTDSEEEIREAFGVFDKDGNGYISAAELRHV MTNLGEKLTDEEVDEMIREADSDGDGQVNYEEFVQMMTAK" CDS 2332..2337 /codon_start=1 /label=linker /note="color: #c0c0c0" /translation="TG" mat_peptide 2338..2394 /label=P2A, cleaved leaving the last pro on the right /note="color: #cc99b2" CDS 2395..2406 /codon_start=1 /label=linker /note="color: #c0c0c0" /translation="GSTA" CDS 2407..2502 /codon_start=1 /label=#1 COX8 sig pept /note="color: #993366" /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP" CDS 2503..2598 /codon_start=1 /label=#2 COX8 sig pept /note="color: #993366" /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP" CDS 2599..2694 /codon_start=1 /label=#3 COX8 sig pept /note="color: #993366" /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP" CDS 2695..2790 /codon_start=1 /label=#4 COX8 sig pept /note="color: #993366" /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP" CDS 2791..2817 /codon_start=1 /label=linker /note="color: #c0c0c0" /translation="RSGSAKDPT" CDS 2818..4167 /codon_start=1 /label=GCaMP6f /note="color: #339966" /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" CDS 2827..2844 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /note="color: #cc99b2" /translation="HHHHHH" CDS 2884..2907 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /note="This feature has 2 segments: 1: 2884 .. 2892 / #cc99b2 2: 2893 .. 2907 / #cc99b2 / enterokinase site Cleavage site after base 2907" /translation="DLYDDDDK" primer_bind complement(4253..4270) /label=BGH rev primer /note="color: black; sequence: TAGAAGGCACAGTCGAGG" terminator 4256..4483 /label=bGH_PA_terminator /note="color: #ffffff" rep_origin 4546..4852 /label=f1_origin /note="color: #ffff00" primer_bind complement(4966..4986) /label=pBABE_3_primer /note="color: black; sequence: ACCCTAACTGACACACATTCC" promoter 5104..5306 /label=SV40_promoter /note="color: #ffffff" primer_bind 5213..5232 /label=SV40pro_F_primer /note="color: black; sequence: TATTTATGCAGAGGCCGAGG" CDS 5358..6158 /codon_start=1 /label=Neo(R) /note="color: #993366" /translation="SFRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" terminator 6340..6456 /label=SV40_PA_terminator /note="color: #ffffff" primer_bind 6426..6445 /label=EBV_rev_primer /note="color: black; sequence: GTGGTTTGTCCAAACTCATC" primer_bind complement(6501..6519) /label=M13_reverse_primer /note="color: black; sequence: GGAAACAGCTATGACCATG" primer_bind complement(6518..6540) /label=M13_pUC_rev_primer /note="color: black; sequence: AGCGGATAACAATTTCACACAGG" rep_origin complement(6892..7511) /direction=LEFT /label=pBR322_origin /note="color: #ffff00" CDS complement(7666..8526) /codon_start=1 /label=Amp(R) /note="color: #ccffcc" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8568..8596) /label=AmpR_promoter /note="color: #ffffff; direction: LEFT" 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 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 accgccatgg tgggaagaaa ctctgctatc gccgctggcg tttgtggcgc cctgttcatc 961 ggctactgca tctacttcga cagaaagagg cggagcgacc ccaacaccgg ttctcaccac 1021 catcaccatc acggcatggc ctctatgaca ggcggacagc agatgggcag agatctgtac 1081 gacgacgatg acaaggacct ggccacaatg gtggacagca gcaggcggaa gtggaacaaa 1141 tggggacacg ccgtcagagc catcggcaga ttgtctagcg ccaacaacac cgagatgatg 1201 taccctgctg acggcggcct gagaggctac acacatatgg ccctgaaagt ggacggcgga 1261 ggccacctgt cttgtagctt cgtgaccacc tacaggtcca agaaaaccgt gggcaacatc 1321 aagatgcccg ccatccacta cgtgtcccac aggctggaaa gactggaaga gtccgacaac 1381 gagatgttcg tggtgcagag ggaacacgcc gtggccaagt ttgttggact cggaggtggc 1441 ggaggaaccg gcggatctat gaacagcctg atcaaagaaa acatgcggat gaaggtggtg 1501 ctggaaggca gcgtgaacgg ccaccagttc aagtgtacag gcgaaggcga gggcaacccc 1561 tacatgggca cacagaccat gaggatcaaa gtgatcgaag gcggccctct gcctttcgct 1621 ttcgacatcc tggccaccag ctttatgtac ggcagcagga ccttcatcaa gtaccccaag 1681 ggaatccccg acttcttcaa gcagagcttc cccgagggct tcacctggga gagagtgacc 1741 agatacgagg acggcggcgt gatcacagtg atgcaggaca cctctctgga agatggctgc 1801 ctggtgtacc acgtgcaagt gcggggagtg aacttccctt ctaacggggc cgtgatgcag 1861 aaaaagacca aaggctggga gcccaccgac agccagctga cagaagaaca gatcgccgag 1921 ttcaaagagg ccttcagcct gttcgacaag gacggcgacg gcacaatcac caccaaagaa 1981 atgggcaccg tgatgagaag cctgggacag aaccctacag aggccgagct gcaggacatg 2041 atcaacgagg ttgacgccga tggcgacgga actatcgact tccctgagtt cctgatcatg 2101 atggccggca agatgaagta caccgactcc gaggaagaga tccgcgaggc cttcggagtg 2161 tttgataagg acggaaacgg ctacatctct gccgccgagc tgagacacgt gatgaccaat 2221 ctgggcgaga agctgaccga cgaagaggtg gacgagatga tcagagaggc cgattcagac 2281 ggcgacgggc aagtgaacta cgaagagttc gtgcagatga tgaccgccaa gaccggtgcc 2341 accaacttta gcctgctgaa acaggccggc gacgtggaag agaatcctgg acctggatcc 2401 accgccatga gcgtgctgac acctctgctg ctgagaggcc tgacaggctc tgctagaagg 2461 ctgcctgtgc ctagagccaa gatccactct ctgggcgacc ctatgtctgt gctgactcca 2521 ctgctcctgc ggggactgac tggatctgct cgtagactgc cagtgccacg ggctaagatt 2581 cacagcctgg gcgatccaat gagtgtgctt accccactcc tgctcagagg actgaccgga 2641 agtgctagac gcctgccagt tccaagggcc aaaatccata gtctgggaga tcccatgtcc 2701 gtgttgacac ccctcttgct gaggggactt accggctcag ctcggagact ccctgttcca 2761 agagcaaaga ttcattccct cggcgaccca agatctggca gcgctaaaga ccctaccatg 2821 ggtagccacc accaccatca ccacggcatg gcttctatga caggcggcca gcagatgggc 2881 cgcgacctgt acgatgacga cgataaggac ctggccacaa tggtggacag cagcaggcgg 2941 aagtggaaca agacaggcca cgccgttaga gccatcggca gactgtctag cctggaaaac 3001 gtgtacatca aggccgacaa gcagaagaac ggcatcaagg ctaacttcaa gatccggcac 3061 aacatcgagg acggcggagt gcagctggct taccactacc agcagaacac ccctatcggc 3121 gacggaccag ttctgctgcc tgacaaccac tacctgagcg tgcagagcaa gctgagcaag 3181 gaccccaacg agaagaggga ccacatggtg ctgctggaat ttgtgaccgc cgctggcatc 3241 acactcggca tggacgaact gtacaaaggc ggcacaggcg gctctatggt gtccaagggc 3301 gaagaactgt tcaccggcgt ggtgcccatt ctggtggaac tggatgggga tgtgaacggc 3361 cacaagttca gcgtcagcgg agaaggcgaa ggcgacgcca catacggaaa gctgaccctg 3421 aagttcatct gcaccaccgg caaactgccc gtgccttggc ctacactggt caccacactg 3481 acatacggcg tgcagtgctt cagcagatac cccgaccata tgaagcagca cgacttcttc 3541 aagagcgcca tgcctgaggg ctacatccaa gagagaacca tcttctttaa ggacgacggc 3601 aactacaaga ccagggccga agtgaagttc gagggcgaca ccctggtcaa cagaatcgag 3661 ctgaagggca tcgacttcaa agaggacggc aacatcctgg gccacaagct cgagtacaac 3721 ctgcctgacc agctgaccga ggaacagatc gccgagttca aagaagagtt cagcctgttc 3781 gacaaggacg gcgacggcac aatcaccacc aaagaactgg gcaccgtgat gagaagcctg 3841 ggacagaacc ctacagaggc cgagctgcag gacatgatca acgaggttga cgctgatggc 3901 gacggaacta tcgacttccc cgagttcctg accatgatgg ccagaaagat gaagtacagg 3961 gacacagagg aagagatccg cgaggccttc ggcgtgttcg ataaggatgg aaacggctac 4021 atctctgccg ctgagctgag acacgtgatg accaacctgg gcgagaagct gacagacgaa 4081 gaggtggacg agatgatcag agaggccgac attgacggcg acgggcaagt gaactacgaa 4141 gagtttgtcc agatgatgac cgccaagtga accggtgtcg acgaattctg cagatatcca 4201 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact 4261 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 4321 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 4381 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 4441 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 4501 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 4561 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 4621 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 4681 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 4741 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 4801 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 4861 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 4921 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 4981 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 5041 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 5101 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 5161 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 5221 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 5281 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 5341 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 5401 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 5461 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 5521 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 5581 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 5641 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 5701 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 5761 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 5821 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 5881 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 5941 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 6001 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 6061 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 6121 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 6181 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 6241 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 6301 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 6361 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 6421 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 6481 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6541 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6661 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6721 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7321 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 7381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8521 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8581 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8641 ccgaaaagtg ccacctgacg tc //