LOCUS       sequence-462146-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector expressing the mScarlet based calcium indicator 
            PinkyCaMP under gfaABC1D promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  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 5898)
  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: pZac2.1-gfaABC1D-PinkyCaMP
FEATURES             Location/Qualifiers
     source          1..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1523..1540
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1544..1576
                     /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(1580..1588,1589..1603)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1646..1702
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2389..2408
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2438..2881
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     polyA_signal    2923..3044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2934..2953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2988..3007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3206..3335
                     /label=AAV2 ITR
     primer_bind     complement(3354..3371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3354..3370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3363..3385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3512..3967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3599..3618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3809..3830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3993..4097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4098..4166,4167..4958)
                     /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"
     rep_origin      5129..5717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5618..5637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     1441 tcgagaattc gccaccatgc tgcagaacga gcttgctctt aagttggctg gacttgatat
     1501 taacaagact ggaggaggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg
     1561 tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatctcg caacaatggt
     1621 cgactcatca atggtcgact catcacgtcg taagtggaat aaggcaggtc acgcagtcag
     1681 agctataggt cggctgagct caagatcttg gtctgtggag cggttgtacc ccgaggacgg
     1741 cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc
     1801 ggacttcaag accacctata ttgccaagaa gcccgtgcag atgcccggcg cctacaacgt
     1861 cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga
     1921 acgctccgtg gcgcgccact ccaccggcgg catggacgag ctgtacaagg gaggaactgg
     1981 tggtagtatg gtgagcaagg gcgaggcagt gatcaaggag ttcatgcgct acaaggtgca
     2041 catggagggc tccgtaaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2101 ctacgagggt acccagaccg ccaagctgac ggtgaccaag ggtggccccc tgcccttctc
     2161 ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcacta agcaccccgc
     2221 cgacatcccc gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2281 gaacttcgag gacggcggcg ccgtggttgt gtttcaggac acctccctgg aggacggcgt
     2341 attcatctac aaggtgaagc tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     2401 gaagaagaca atgggctgga tgccggcgtt tattctggac caactgactg aagagcagat
     2461 cgcagaattt aaagaggctt tctccctatt tgacaaggac ggggatggga cgataacaac
     2521 caaggagctg gggacggtga tgcggtctct ggggcagaac cccacagaag cagagctgca
     2581 ggacatgatc aatgaagtag atgccgacgg tgacggcaca ttcgacttcc ctgagttcct
     2641 gacgatgatg gcaagaaaaa tgaatgacac agacagtgaa gaggaaatta gagaagcgtt
     2701 ccgcgtgttt gataaggacg gcaatggcta catcggcgca gcagagcttc gccacgtgat
     2761 gacagacctt ggagagaagt taacagatga ggaggttgat gaaatgatca gggtagcaga
     2821 catcgatggg gatggtcagg taaactacga agagtttgta caaatgatga cagcgaagtg
     2881 aattctagag tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     2941 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3001 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3061 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3121 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     3181 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     3241 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3301 gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc
     3361 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3421 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3481 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3541 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3601 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3661 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3721 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3781 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3841 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3901 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3961 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4021 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4081 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4141 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4201 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4261 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4321 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4381 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4441 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4501 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4561 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4621 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4681 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4741 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4801 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4861 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4921 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4981 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5041 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5881 tgagcgagga agcggaag
//
