LOCUS       sequence-291768-        6683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CaMPARI V398D in mammalian cells, a photoconvertible 
            fluorescent protein-based calcium integrator.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2015-02-13- 13"; volume: "347"; 
            issue: "6223"; pages: "755-60"
COMMENT     Sequence Label: pcDNA3-CaMPARI V398D
FEATURES             Location/Qualifiers
     source          1..6683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             964..981
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             988..1428
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2149..2205
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHADRAIGRLSS"
     promoter        complement(2236..2254)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2237..2254)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2274..2291)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2280..2504
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2550..2978
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2637..2656)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2847..2868
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2987..3007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2992..3321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3172..3307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3234..3253
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3388..4182
                     /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(3442..3461)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4052..4071
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4356..4477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4393..4412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4447..4466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4526..4542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4526..4542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4539..4561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4550..4566
                     /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(join(4574..4580,4581..4598,4599..4604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4619..4640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4757..4774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4928..5516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5008..5027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5687..6478,6479..6547))
                     /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     6310..6329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6548..6652)
                     /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 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atgctgcaga acgagcttgc tcttaagttg gctggacttg atattaacaa gactggaggt
      961 tctcatcatc accaccacca tggatccgac caactgactg aagagcagat cgcagaattt
     1021 aaagaggctt tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg
     1081 gggacggtga tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc
     1141 aatgaagtag atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg
     1201 gcaagaaaaa tgaaagacac agacagtgaa gaagaaatta gagaagcgtt ccgtgtgttt
     1261 gataaggatg gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt
     1321 ggagagaagt taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg
     1381 gatggtcagg taaactacga agagtttgta caaatgatga cagcgaagct cgagtgcgag
     1441 aaaatctatg tgcgtgatgg agtgctgacg ggtgatattc atatgatctt gttgcttgaa
     1501 ggaaatgccc attaccgatg tgacttcaga actacttaca aagctaagga gaagggtgtc
     1561 aagttaccag gcgtgcactt tgtggaccac tgcattgaga ttttaagcca tgacaaagat
     1621 tacaacaagg ttaagctgta tgagtatgct gttgctcatt ctggattgcc tgacaatgcc
     1681 agacgaggcg gtaccggcgg atccatggtg agtgcgatta agccagacat gaagatcaaa
     1741 ctccgtatgg aaggcaacgt aaacgggcac cactttgtga tcgacggaga tggtacaggc
     1801 aagccttatg agggaaaaca gaccatggat cttgaagtca aagagggcgg acctctgcct
     1861 tttgcctttg atatcctgac cactgcattc cattacggca acagggtatt cgtgaaatat
     1921 ccagacaaca tacaagacta ttttaagcag tcgtttccta aggggtattc gtgggaacga
     1981 agcatgactt tcgaagacgg gggcatttgc tatgccagaa acgacataac aatggaaggg
     2041 gacactttct ataataaagt tcgattttat ggtaccaact ttcccgccaa tggtccagtt
     2101 atgcagaaga agacgctgaa atggatgccg agctggacgc gttcatcacg tcgtaagtgg
     2161 aataagacag gtcacgcaga cagagctata ggtcggctga gctcataact cgagcatgca
     2221 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     2281 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2341 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2401 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2461 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2521 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2581 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2641 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2701 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2761 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2821 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2881 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2941 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3001 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3061 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3121 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3241 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3301 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3361 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3421 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3481 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3541 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3601 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3661 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3721 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3781 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3841 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3901 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3961 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4021 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4081 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4141 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4201 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4261 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4321 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4381 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4441 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4501 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4561 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4621 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4681 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4741 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4801 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4861 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4921 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4981 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5041 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5101 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5161 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5221 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5281 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5341 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5401 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5461 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5581 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5641 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5701 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5761 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5821 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5881 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5941 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6001 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6061 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6121 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6181 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6241 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6301 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6361 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6421 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6481 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6541 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6601 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6661 cccgaaaagt gccacctgac gtc
//
