LOCUS       sequence-462152-        6780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector expressing the mScarlet based calcium indicator 
            PinkyCaMP under CaMKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6780)
  AUTHORS   Fink R, Imai S, Gockel N, Lauer G, Renken K, Wietek J, 
            Lamothe-Molina PJ, Furhmann F, Mittag M, Ziebarth T, Canziani A, 
            Kubitschke M, Kistmacher V, Kretschmer A, Sebastian E, Schmitz D, 
            Terai T, Grundemann J, Hassan S, Patriarchi T, Reiner A, Fuhrmann M,
            Campbell RE, Masseck OA
  TITLE     PinkyCaMP a mScarlet-based calcium sensor with exceptional 
            brightness, photostability, and multiplexing capabilities.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 11:2024.12.16.628673. doi: 
            10.1101/2024.12.16.628673.
  PUBMED    39763884
REFERENCE   2  (bases 1 to 6780)
  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"; journalName: "bioRxiv 
            [Preprint]. 2025 Jan 11:2024.12.16.628673. doi: 
            10.1101/2024.12.16.628673."
COMMENT     Sequence Label: pAAV-CaMKII-PinkyCaMP
FEATURES             Location/Qualifiers
     source          1..6780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             1524..1541
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1545..1577
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1581..1589,1590..1604)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1647..1703
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2390..2409
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2439..2882
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     misc_feature    2907..3495
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2960..2980)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3378..3389)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3527..4003
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3726..3745)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4043..4183
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4043..4172
                     /label=AAV2 ITR
     rep_origin      4258..4713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4345..4364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4555..4576
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4730..4749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4849..4871
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4909..4927)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4995..5099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5100..5168,5169..5960)
                     /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     complement(5318..5337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6131..6719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6620..6639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccatg ctgcagaacg agcttgctct taagttggct ggacttgata
     1501 ttaacaagac tggaggaggt tctcatcatc atcatcatca tggtatggct agcatgactg
     1561 gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatctc gcaacaatgg
     1621 tcgactcatc aatggtcgac tcatcacgtc gtaagtggaa taaggcaggt cacgcagtca
     1681 gagctatagg tcggctgagc tcaagatctt ggtctgtgga gcggttgtac cccgaggacg
     1741 gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg
     1801 cggacttcaa gaccacctat attgccaaga agcccgtgca gatgcccggc gcctacaacg
     1861 tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg
     1921 aacgctccgt ggcgcgccac tccaccggcg gcatggacga gctgtacaag ggaggaactg
     1981 gtggtagtat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgc tacaaggtgc
     2041 acatggaggg ctccgtaaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2101 cctacgaggg tacccagacc gccaagctga cggtgaccaa gggtggcccc ctgcccttct
     2161 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcact aagcaccccg
     2221 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     2281 tgaacttcga ggacggcggc gccgtggttg tgtttcagga cacctccctg gaggacggcg
     2341 tattcatcta caaggtgaag ctgcgcggca ccaacttccc ccccgacggc cccgtaatgc
     2401 agaagaagac aatgggctgg atgccggcgt ttattctgga ccaactgact gaagagcaga
     2461 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acgataacaa
     2521 ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc
     2581 aggacatgat caatgaagta gatgccgacg gtgacggcac attcgacttc cctgagttcc
     2641 tgacgatgat ggcaagaaaa atgaatgaca cagacagtga agaggaaatt agagaagcgt
     2701 tccgcgtgtt tgataaggac ggcaatggct acatcggcgc agcagagctt cgccacgtga
     2761 tgacagacct tggagagaag ttaacagatg aggaggttga tgaaatgatc agggtagcag
     2821 acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt
     2881 gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2941 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3001 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3061 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3121 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3181 ccggaacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3241 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3301 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3361 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3421 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3481 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3541 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3601 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3661 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3721 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3781 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3841 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3901 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3961 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4021 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4081 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4141 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4201 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4261 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4321 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4381 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4441 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4501 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4561 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4621 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4681 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4741 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4801 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4861 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4921 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4981 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5041 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5101 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5161 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5221 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5281 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5341 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5401 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5461 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5521 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5581 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5641 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5701 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5761 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5821 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5881 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5941 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6001 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6061 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
