LOCUS       sequence-275078-        6663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytoskeleton, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6663)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 6663)
  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"
COMMENT     Sequence Label: mApple-Myotilin-C-14
FEATURES             Location/Qualifiers
     source          1..6663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    130..177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     202..221
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(409..1203)
                     /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             complement(409..1203)
                     /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(520..539)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1130..1149
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1238..1434)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(1246..1456)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1252..1387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1480..1500
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1525..1629)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1656..2111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1743..1762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1953..1974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2118..2239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2129..2148)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2183..2202
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3905..4615,4616..4618,4619..4621))
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3947..3968)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4296..4315
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4555..4576
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4618..4627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4635..5342)
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4910..4929)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      5339..5348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5387..5590)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5416..5436)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     rep_origin      complement(5876..6464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5956..5975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
       61 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
      121 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
      181 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
      241 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
      301 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
      361 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
      421 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
      481 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
      541 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
      601 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
      661 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
      721 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
      781 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
      841 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
      901 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
      961 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     1021 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     1081 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     1141 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     1201 catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa
     1261 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     1321 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     1381 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     1441 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     1501 acagctggtt ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca
     1561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1621 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcactaag
     1681 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1741 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1801 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1861 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1921 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1981 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2041 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2101 gcttacaatt tacgccttaa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2161 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2221 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2281 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     2341 tgatcagtta tctagatccg gtggatcctt aaagttcttc actttcatag agtccagatt
     2401 gagctgccaa acgctgaaat tctccttctg ggttaaaagc ttgttttaca ttcaaacctt
     2461 tcccattaag tgctaaatat ttgctgaatg ttggtcgaac ccgtaactgc ttaggagctg
     2521 gaagagtttg gtttggacgt gccgtaacgt ctaatcttgt gttacatgta gtcactccag
     2581 cttcattaac tgctgacaca gtataccacc cagcatcttt cttgtttaca tcttttatca
     2641 gtaaagtaac tcttccagtg ttatcttgat ataagcttat tcggtcagtg ttgaattgta
     2701 ccatttcatt atttcttttc cagaaaagct ttggtggagg tatagccgag atctggcatt
     2761 ctagtttcac tgaatctccc tctaaaactt ttttgctctg tggtttgtag ataaacattg
     2821 gtgctctttt atgttctttt gcaaggacat ccagctgcac agtgaaggtg gcttctcctg
     2881 ctctattctt ggcaacacat gcataagccc ctgcatctga agctctgact acttcaaaga
     2941 tgagtgaatg aagacccttc tcagacacta tcattttgtg caaatcatct gattgaactg
     3001 ttcttccatt tagataccat gacacatcag gagctggcag tccactcact ttgaagtcca
     3061 ttctgcagaa tcttccttca tcaatcgaca tgttctctgg cacttgaatg aaacgtggtg
     3121 ggtaaaattt ctcctggatt gcatcctgat cattcacatc tccccttgag gttgatctac
     3181 ttcttacttg tgatgtagga acttgcagtc gcgcatgttc tgagttgtgt tgctgcgagt
     3241 cttgtgcacc actgtcatcc tgagcttgaa acacagcagc tgcattctgt ggtcctagca
     3301 atctgcgagc catcttctct tcatatgtta gacgttgatt tccattatga agaagggtgt
     3361 ccttgcattt cagctttctt tccaaatctt gaatcaaagc ttcttttgat ccttgtattt
     3421 catgatcagg agttcttggt atgggcttag catttgcagt ttgggatggc tttgcattta
     3481 taggttggcc agctgaggac tgttgatagt tggaatccat agcggaaggg attttactgc
     3541 tattgtaatc aggctgtgat ggtaatatgg agctgaggaa gctggctggg gactggttat
     3601 aggtggttgt aaccctttgg cctgggttgg agccagcatg ctgctgggga gaagcaggga
     3661 aagcagagga ggacatggtg atgtgagagc tcagtgttga ggaggcagaa aatctttgct
     3721 ctgtacactg gcggggctgg atgataatgg aagactgttt ggtctggcta gagaagctgg
     3781 aggtttctgg tccaggaggc tgcaatctgg agccacatgg gttttgggac tggatgaagt
     3841 gttttggacg ttcgtagtta aaaccggagc ctccgcttcc gcttgcgctc ccagcacttc
     3901 cggacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     3961 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg gtgctcaggt
     4021 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     4081 cggcgagctg cacgctgccg tcctcgatgt tgtggcgggt cttgaagttc accttgatgc
     4141 cgttcttctg cttgtcggcg gtgatataga ccttgtggct gttgtagttg tactccagct
     4201 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     4261 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     4321 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     4381 gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtaggtcaag gtggtcacga
     4441 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     4501 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     4561 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     4621 tggtggcgac cggacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg
     4681 cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg acacgatgtc caacttgatg
     4741 tcgacgatgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc
     4801 tcggcggcgt agtggccgcc gtccttcagc ttcagcctct tcttgatctc gctcttcagg
     4861 gcgccgtcct cggggtacat ccgctcctcg gaggcctccc agcccatggt cttcttctgc
     4921 attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaacacg
     4981 ccgtcctgca gggaggagtc ctggttaacg tgaataatgc cgccgtcctc gaagttcatc
     5041 acgcgctccc acctgaagcc ctcggggaag gacagcttga agtagtcggg gatgtcggct
     5101 gggtgcttaa tgtagacctt ggagccgtac atgaactgag gggacaggat gtcccaggcg
     5161 aagggcaggg ggccaccctt ggtcaccttc agcttagcgg tctgaaaggc ctcgtagggg
     5221 cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc
     5281 accttgaagc gcatgaactc cttgatgatg gccatgttat tctcctcgcc cttgctcacc
     5341 atggtggcga ccggtagcgc tagcggatct gacggttcac taaaccagct ctgcttatat
     5401 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     5461 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     5521 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     5581 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     5641 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtaa gttatgtaac
     5701 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     5761 aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5821 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5881 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5941 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6001 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6061 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6121 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6181 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6241 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6301 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6361 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6421 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6481 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6541 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6601 tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg
     6661 cga
//
