LOCUS Exported 6683 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses CaMPARI in mammalian cells, a photoconvertible fluorescent protein-based calcium integrator. ACCESSION . VERSION . KEYWORDS pcDNA3-CaMPARI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6683) AUTHORS Fosque BF, Sun Y, Dana H, Yang CT, Ohyama T, Tadross MR, Patel R, Zlatic M, Kim DS, Ahrens MB, Jayaraman V, Looger LL, Schreiter ER TITLE Neural circuits. Labeling of active neural circuits in vivo with designed calcium integrators. JOURNAL Science. 2015 Feb 13;347(6223):755-60. doi: 10.1126/science.1260922. PUBMED 25678659 REFERENCE 2 (bases 1 to 6683) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6683 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(75..94) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 266..645 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 646..849 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 800..820 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 894..913 /label=T7 /note="T7 promoter, forward primer" promoter 894..912 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 995..1012 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1019..1459 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" CDS 2180..2236 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" promoter complement(2267..2285) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2268..2285) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2305..2322) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2311..2535 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2581..3009 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2668..2687) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2878..2899 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3018..3038) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3023..3352 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3203..3338 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3265..3284 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3419..4213 /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(3473..3492) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4083..4102 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4387..4508 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4424..4443) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4478..4497 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4557..4573) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4557..4573) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4570..4592) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4581..4597 /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(4605..4635) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4650..4671 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4788..4805) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4959..5547) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5039..5058) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5718..6578) /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 6341..6360 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6579..6683) /gene="bla" /label=AmpR promoter ORIGIN 1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta 61 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct 121 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa 181 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc 241 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca 301 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta 361 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat 421 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg 481 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac 541 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt 601 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg 661 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc 721 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt 781 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata 841 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg 901 actcactata gggagaccca agcttgccac catgctgcag aacgagcttg ctcttaagtt 961 ggctggactt gatattaaca agactggagg ttctcatcat caccaccacc atggatccga 1021 ccaactgact gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga 1081 cggggatggg acaataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa 1141 ccccacagaa gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac 1201 aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa atgaaagaca cagacagtga 1261 agaagaaatt agagaagcgt tccgtgtgtt tgataaggat ggcaatggct acatcagtgc 1321 agcagagctt cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga 1381 tgaaatgatc agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt 1441 acaaatgatg acagcgaagc tcgagtgcga gaaaatctat gtgcgtgatg gagtgctgac 1501 gggtgatatt catatgatct tgttgcttga aggaaatgcc cattaccgat gtgacttcag 1561 aactacttac aaagctaagg agaagggtgt caagttacca ggcgtgcact ttgtggacca 1621 ctgcattgag attttaagcc atgacaaaga ttacaacaag gttaagctgt atgagtatgc 1681 tgttgctcat tctggattgc ctgacaatgc cagacgaggc ggtaccggcg gatccatggt 1741 gagtgcgatt aagccagaca tgaagatcaa actccgtatg gaaggcaacg taaacgggca 1801 ccactttgtg atcgacggag atggtacagg caagccttat gagggaaaac agaccatgga 1861 tcttgaagtc aaagagggcg gacctctgcc ttttgccttt gatatcctga ccactgcatt 1921 ccattacggc aacagggtat tcgtgaaata tccagacaac atacaagact attttaagca 1981 gtcgtttcct aaggggtatt cgtgggaacg aagcatgact ttcgaagacg ggggcatttg 2041 ctatgccaga aacgacataa caatggaagg ggacactttc tataataaag ttcgatttta 2101 tggtaccaac tttcccgcca atggtccagt tatgcagaag aagacgctga aatggatgcc 2161 gagctggacg cgttcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat 2221 aggtcggctg agctcataac tcgagcatgc atctagaggg ccctattcta tagtgtcacc 2281 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg 2341 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct 2401 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg 2461 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg 2521 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc 2581 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 2641 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 2701 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 2761 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 2821 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 2881 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 2941 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 3001 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 3121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 3181 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 3241 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 3301 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 3361 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 3421 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 3481 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 3541 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 3601 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 3661 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 3721 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 3781 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 3841 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 3901 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 3961 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 4021 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 4081 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 4141 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 4201 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg 4261 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 4321 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 4381 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4441 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4501 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 4561 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 4621 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 4681 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 4741 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4801 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4861 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 5161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 5401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 5821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 6001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6301 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 6601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 6661 aataaacaaa taggggttcc gcg //