LOCUS       sequence-463043-        5386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A vector expressing the mScarlet based calcium indicator PinkyCaMP 
            under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5386)
  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 5386)
  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: pN1-PinkyCaMP
FEATURES             Location/Qualifiers
     source          1..5386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             694..711
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             715..747
                     /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(751..759,760..774)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             817..873
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1560..1579
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1609..2052
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     polyA_signal    2174..2295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2211..2230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2265..2284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2302..2757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2439..2460)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2651..2670
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2784..2888
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2890..3247
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2890..3081
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2913..2933)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3098..3233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3282..4076
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3282..4076
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3336..3355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3946..3965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4264..4283)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4308..4355
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4684..5272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5173..5192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatg ctgcagaacg agcttgctct taagttggct
      661 ggacttgata ttaacaagac tggaggaggt tctcatcatc atcatcatca tggtatggct
      721 agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatctc
      781 gcaacaatgg tcgactcatc aatggtcgac tcatcacgtc gtaagtggaa taaggcaggt
      841 cacgcagtca gagctatagg tcggctgagc tcaagatctt ggtctgtgga gcggttgtac
      901 cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc
      961 cgctacctgg cggacttcaa gaccacctat attgccaaga agcccgtgca gatgcccggc
     1021 gcctacaacg tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg
     1081 gaacagtacg aacgctccgt ggcgcgccac tccaccggcg gcatggacga gctgtacaag
     1141 ggaggaactg gtggtagtat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgc
     1201 tacaaggtgc acatggaggg ctccgtaaac ggccacgagt tcgagatcga gggcgagggc
     1261 gagggccgcc cctacgaggg tacccagacc gccaagctga cggtgaccaa gggtggcccc
     1321 ctgcccttct cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcact
     1381 aagcaccccg ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg
     1441 gagcgcgtga tgaacttcga ggacggcggc gccgtggttg tgtttcagga cacctccctg
     1501 gaggacggcg tattcatcta caaggtgaag ctgcgcggca ccaacttccc ccccgacggc
     1561 cccgtaatgc agaagaagac aatgggctgg atgccggcgt ttattctgga ccaactgact
     1621 gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg
     1681 acgataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa
     1741 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac attcgacttc
     1801 cctgagttcc tgacgatgat ggcaagaaaa atgaatgaca cagacagtga agaggaaatt
     1861 agagaagcgt tccgcgtgtt tgataaggac ggcaatggct acatcggcgc agcagagctt
     1921 cgccacgtga tgacagacct tggagagaag ttaacagatg aggaggttga tgaaatgatc
     1981 agggtagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg
     2041 acagcgaagt gagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     2101 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2161 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2221 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2281 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2341 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2401 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2461 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2521 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2581 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2641 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2701 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2881 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2941 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3001 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3121 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3181 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3241 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3301 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3361 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3421 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3481 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3541 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3601 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3661 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3721 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3781 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3841 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3901 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3961 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4021 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4081 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4141 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4201 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4261 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4321 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc tgtcccaggg
     4381 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4441 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4501 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4681 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4741 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4801 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4861 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4921 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4981 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5041 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5101 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5161 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5221 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5281 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5341 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
