LOCUS       sequence-307040-        6447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged MyTH/FERM domains of MYO10..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6447)
  AUTHORS   Miihkinen M, Gronloh MLB, Popovic A, Vihinen H, Jokitalo E, Goult 
            BT, Ivaska J, Jacquemet G
  TITLE     Myosin-X and talin modulate integrin activity at filopodia tips.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109716. doi: 
            10.1016/j.celrep.2021.109716.
  PUBMED    34525374
REFERENCE   2  (bases 1 to 6447)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109716. doi"; pages: " 
            10.1016/j.celrep.2021.109716"
COMMENT     Sequence Label: EGFP-FERM (Myo10)
FEATURES             Location/Qualifiers
     source          1..6447
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2028..2037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3235..3356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3272..3291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3326..3345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3363..3818)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3500..3521)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3712..3731
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3845..3949
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3951..4308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3951..4142
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3974..3994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4159..4294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4343..5137
                     /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             4343..5137
                     /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(4397..4416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5007..5026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5325..5344)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5369..5416
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5745..6333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6234..6253
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attcaacccg gtggtccagt
     1381 gccattcaaa acgtgactga caccaaggcc ccgatcgaca cccccaccca gcagctgatt
     1441 caagatatca aggagaactg cctgaactcg gatgtggtgg aacagattta caagcggaac
     1501 ccgatccttc gatacaccca tcaccccttg cactccccgc tcctgcccct tccgtatggg
     1561 gacataaatc tcaacttgct caaagacaaa ggctatacca cccttcagga tgaggccatc
     1621 aagatattca attccctgca gcaactggag tccatgtctg acccaattcc aataatccag
     1681 ggcatcctac agacagggca tgacctgcga cctctgcggg acgagctgta ctgccagctt
     1741 atcaaacaga ccaacaaagt gccccacccc ggcagtgtgg gcaacctgta cagctggcag
     1801 atcctgacat gcctgagctg caccttcctg ccgagtcgag ggattctcaa gtatctcaag
     1861 ttccatctga aaaggatacg ggaacagttt ccaggaaccg agatggaaaa atacgctctc
     1921 ttcacttacg aatctcttaa gaaaaccaaa tgccgagagt ttgtgccttc ccgagatgaa
     1981 atagaagctc tgatccacag gcaggaaatg acatccacgg tctattgcca tggcggcggc
     2041 tcctgcaaga tcaccatcaa ctcccacacc actgctgggg aggtggtgga gaagctgatc
     2101 cgaggcctgg ccatggagga cagcaggaac atgtttgctt tgtttgaata caacggccac
     2161 gtcgacaaag ccattgaaag tcgaaccgtc gtagctgatg tcttagccaa gtttgaaaag
     2221 ctggctgcca catccgaggt tggggacctg ccatggaaat tctacttcaa actttactgc
     2281 ttcctggaca cagacaacgt gccaaaagac agtgtggagt ttgcatttat gtttgaacag
     2341 gcccacgaag cggttatcca tggccaccat ccagccccgg aagaaaacct ccaggttctt
     2401 gctgccctgc gactccagta tctgcagggg gattatactc tgcacgctgc catcccacct
     2461 ctcgaagagg tttattccct gcagagactc aaggcccgca tcagccagtc aaccaaaacc
     2521 ttcacccctt gtgaacggct ggagaagagg cggacgagct tcctagaggg gaccctgagg
     2581 cggagcttcc ggacaggatc cgtggtccgg cagaaggtcg aggaggagca gatgctggac
     2641 atgtggatta aggaagaagt ctcctctgct cgagccagta tcattgacaa gtggaggaaa
     2701 tttcagggaa tgaaccagga acaggccatg gccaagtaca tggccttgat caaggagtgg
     2761 cctggctatg gctcgacgct gtttgatgtg gagtgcaagg aaggtggctt ccctcaggaa
     2821 ctctggttgg gtgtcagcgc ggacgccgtc tccgtctaca agcgtggaga gggaagacca
     2881 ctggaagtct tccagtatga acacatcctc tcttttgggg cacccctggc gaatacgtat
     2941 aagatcgtgg tcgatgagag ggagctgctc tttgaaacca gtgaggtggt ggatgtggcc
     3001 aagctcatga aagcctacat cagcatgatc gtgaagaagc gctacagcac gacacgctcc
     3061 gccagcagcc agggcagctc caggtgataa ggtaccgcgg gcccgggatc caccggatct
     3121 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3181 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3241 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3301 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3361 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3421 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3481 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3541 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3601 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3661 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3721 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3781 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3841 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3901 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3961 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4141 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4261 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4321 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4381 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4441 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4501 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4561 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4621 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4681 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4741 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4801 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4861 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4921 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4981 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5041 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5101 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5161 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5221 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5281 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5341 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5401 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5461 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5521 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5581 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5641 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5701 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5761 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5821 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5881 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5941 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6001 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6061 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6121 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6181 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6241 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6301 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6361 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6421 gtggataacc gtattaccgc catgcat
//
