LOCUS       sequence-247900-        5266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a fusion between myristoylation signal peptide and 
            R-GECO1. This construct targets R-GECO1 to the plasma membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5266)
  AUTHORS   Kim JM, Lee M, Kim N, Heo WD
  TITLE     Optogenetic toolkit reveals the role of Ca2+ sparklets in 
            coordinated cell migration.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 24;113(21):5952-7. doi: 
            10.1073/pnas.1518412113. Epub 2016 May 17.
  PUBMED    27190091
REFERENCE   2  (bases 1 to 5266)
  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: 
            "10.1073/pnas.1518412113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-24- 24"; volume: "113"; issue: "21"; pages: "5952-7"
COMMENT     Sequence Label: Lyn-R-GECO1
FEATURES             Location/Qualifiers
     source          1..5266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             634..690
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1386..1405
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1429..1872
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     polyA_signal    1994..2115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2031..2050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2085..2104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2122..2577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2259..2280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2471..2490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2604..2708
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2710..3067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2710..2901
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2733..2753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2918..3053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3102..3896
                     /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             3102..3896
                     /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(3156..3175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3766..3785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4084..4103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4128..4175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4504..5092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4993..5012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
      541 atgggatgta taaaatcaaa agggaaagac agcgcgggag cagatagtgc tggtagtgct
      601 ggtagtgctg ggaattcgat ggtcgactca tcacgtcgta agtggaataa ggcaggtcac
      661 gcagtcagag ctataggtcg gctgagctca cccgtggttt ccgagcggat gtaccccgag
      721 gacggcgccc tgaagagcga gatcaagaag gggctgaggc tgaaggacgg cggccactac
      781 gccgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
      841 atcgtcgaca tcaagttgga catcgtgtcc cacaacgagg actacaccat cgtggaacag
      901 tgcgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggaggt
      961 acaggcggga gtctggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1021 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1081 gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt
     1141 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1201 tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc
     1261 aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta ttcacgttaa ccaggactcc
     1321 tccctgcagg acggcgtatt catctacaag gtgaagctgc gcggcaccaa cttccccccc
     1381 gacggccccg taatgcagaa gaagaccatg ggctgggagg ctacgcgtga ccaactgact
     1441 gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg
     1501 acgataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa
     1561 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac attcgacttc
     1621 cctgagttcc tgacgatgat ggcaagaaaa atgaatgaca cagacagtga agaggaaatt
     1681 agagaagcgt tccgcgtgtt tgataaggac ggcaatggct acatcggcgc agcagagctt
     1741 cgccacgtga tgacagacct tggagagaag ttaacagatg aggaggttga tgaaatgatc
     1801 agggtagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg
     1861 acagcgaagt aggcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1921 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1981 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2101 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2161 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2221 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2281 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2341 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2401 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2461 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2521 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2641 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2701 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2761 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2821 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2941 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3001 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3061 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3601 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4081 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4141 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4201 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4261 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4321 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4381 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4501 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4561 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4621 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4681 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4741 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4801 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4861 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4921 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4981 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5041 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5101 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5161 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     5221 aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
