LOCUS       sequence-97764-e        6684 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 6684)
  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 6684)
  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..6684
                     /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..3322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3173..3308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3235..3254
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3389..4183
                     /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(3443..3462)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4053..4072
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4357..4478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4394..4413)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4448..4467
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4527..4543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4527..4543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4540..4562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4551..4567
                     /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(4575..4581,4582..4599,4600..4605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4620..4641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4758..4775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4929..5517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5009..5028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5688..6479,6480..6548))
                     /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     6311..6330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6549..6653)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2761 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2821 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2881 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     2941 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3001 ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct
     3061 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3121 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3241 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3301 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3361 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3421 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3481 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3541 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3601 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3661 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3721 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3781 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3841 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3901 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3961 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4021 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4081 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4141 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4201 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4261 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4321 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4381 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4441 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4501 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4561 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4621 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4681 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4741 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4801 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4861 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4921 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4981 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5041 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5101 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5161 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5221 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5281 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5341 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5401 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5461 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5521 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5581 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5641 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5701 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5761 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5821 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5881 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5941 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6001 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6061 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6121 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6181 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6241 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6301 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6361 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6421 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6481 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6541 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6601 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6661 ccccgaaaag tgccacctga cgtc
//
