LOCUS       sequence-463044-        6133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector expressing the mScarlet based calcium indicator 
            PinkyCaMP under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6133)
  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 6133)
  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-CAG-PinkyCaMP
FEATURES             Location/Qualifiers
     source          1..6133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   162..291
                     /label=AAV2 ITR
     primer_bind     1169..1191
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1253..1272
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1377..1394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1398..1430
                     /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(1434..1442,1443..1457)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1500..1556
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2243..2262
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2292..2735
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     misc_feature    2760..3348
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2813..2833)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3231..3242)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3391..3512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3402..3421)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3456..3475
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4018..4606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4098..4117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4777..5568,5569..5637))
                     /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     5400..5419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5638..5742)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5768..6133,1..90))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5905..5926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6117..6133,1..3)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
       61 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
      121 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca
      181 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
      241 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
      301 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagagt accattgacg
      361 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      421 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      481 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      541 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      601 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
      661 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
      721 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
      781 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
      841 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      901 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      961 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
     1021 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
     1081 ggctccggga gggccctttg tgcgggggga gcggctcggg gctgtccgcg gggggacggc
     1141 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1201 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1261 ctggttattg tgctgtctca tcattttggc aaagaattgg atccgccacc atgctgcaga
     1321 acgagcttgc tcttaagttg gctggacttg atattaacaa gactggagga ggttctcatc
     1381 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
     1441 acgacgatga cgataaggat ctcgcaacaa tggtcgactc atcaatggtc gactcatcac
     1501 gtcgtaagtg gaataaggca ggtcacgcag tcagagctat aggtcggctg agctcaagat
     1561 cttggtctgt ggagcggttg taccccgagg acggcgtgct gaagggcgac attaagatgg
     1621 ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc tatattgcca
     1681 agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac atcacctccc
     1741 acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgtggcgcgc cactccaccg
     1801 gcggcatgga cgagctgtac aagggaggaa ctggtggtag tatggtgagc aagggcgagg
     1861 cagtgatcaa ggagttcatg cgctacaagg tgcacatgga gggctccgta aacggccacg
     1921 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggtacccag accgccaagc
     1981 tgacggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
     2041 tgtacggctc cagggccttc actaagcacc ccgccgacat ccccgactac tataagcagt
     2101 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtgg
     2161 ttgtgtttca ggacacctcc ctggaggacg gcgtattcat ctacaaggtg aagctgcgcg
     2221 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gacaatgggc tggatgccgg
     2281 cgtttattct ggaccaactg actgaagagc agatcgcaga atttaaagag gctttctccc
     2341 tatttgacaa ggacggggat gggacgataa caaccaagga gctggggacg gtgatgcggt
     2401 ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg
     2461 acggtgacgg cacattcgac ttccctgagt tcctgacgat gatggcaaga aaaatgaatg
     2521 acacagacag tgaagaggaa attagagaag cgttccgcgt gtttgataag gacggcaatg
     2581 gctacatcgg cgcagcagag cttcgccacg tgatgacaga ccttggagag aagttaacag
     2641 atgaggaggt tgatgaaatg atcagggtag cagacatcga tggggatggt caggtaaact
     2701 acgaagagtt tgtacaaatg atgacagcga agtgagaatt cgatatcaag cttatcgata
     2761 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2821 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2881 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2941 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3001 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3061 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3121 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3181 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3241 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3301 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3361 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3421 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3481 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3541 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3601 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3661 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3721 tcgctcactg aggccgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3781 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5281 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5341 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5401 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5461 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5521 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5581 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5761 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5821 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5881 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5941 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6001 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6061 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6121 aagaaagcga aag
//
