LOCUS       sequence-310127-        4841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  F-actin bidning mutant of MPAct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4841)
  AUTHORS   Bisaria A, Hayer A, Garbett D, Cohen D, Meyer T
  TITLE     Membrane-proximal F-actin restricts local membrane protrusions and 
            directs cell migration.
  JOURNAL   Science. 2020 Jun 12;368(6496):1205-1210. doi: 
            10.1126/science.aay7794.
  PUBMED    32527825
REFERENCE   2  (bases 1 to 4841)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2020-06-12- 12"; volume: "368"; 
            issue: "6496"; pages: "1205-1210"
COMMENT     Sequence Label: C1-MPAct(delta1-6)-mRuby3
FEATURES             Location/Qualifiers
     source          1..4841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     187..206
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(505..524)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    549..596
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      925..1513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1414..1433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1688..1991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1992..2195
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2146..2166
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2348..3058
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     polyA_signal    3256..3377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3293..3312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3347..3366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3384..3839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3521..3542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3733..3752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3866..3970
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3972..4329
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3972..4163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3995..4015)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4180..4315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4364..4841,1..317)
                     /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             join(4364..4841,1..317)
                     /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(4418..4437)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
       61 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
      121 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
      181 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
      241 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
      301 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
      361 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
      421 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
      481 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
      541 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
      601 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
      661 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
      721 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
      781 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
      841 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      901 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      961 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1021 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1081 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1141 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1201 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1261 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1321 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1381 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1441 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1501 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1561 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1621 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     1681 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     1741 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     1801 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     1861 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1921 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1981 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     2041 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     2101 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     2161 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
     2221 agcgctaccg gactcagatc tcgagctcaa gcttatgggc atggcgcgac cacggggcgc
     2281 ggggccctgc agccccgggt tggagcgggc tccgcgccgg agcgtcgggg agtcggatcc
     2341 accggtcatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt
     2401 catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcagacc
     2461 gtacgaggga gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc
     2521 ctttgacatt cttgccacgt cgttcatgta tggcagccgt acctttatca agtacccggc
     2581 cgacatccct gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac
     2641 gagatacgaa gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga
     2701 gctcgtctac aacgtcaagg tcagaggggt aaactttccc tccaatggtc ccgtgatgca
     2761 gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct
     2821 gagaggatac actgacatcg cactgaaagt tgatggtggt ggccatctgc actgcaactt
     2881 cgtgacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc
     2941 cgttgatcac cgcctggaaa ggatcgagga gagtgacaat gaaacctacg tagtgcaaag
     3001 agaagtggca gttgccaaat acagcaacct tggtggtggc atggacgagc tgtacaagag
     3061 atctcgagct caagcttcga attctggtaa aaagaagaaa aagaagtcaa agacaaagtg
     3121 tgtaattatg tagagcggcc gcgactctag atcataatca gccataccac atttgtagag
     3181 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3241 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3361 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3421 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3481 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3541 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3601 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3661 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3721 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3781 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3961 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4021 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4081 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4201 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4261 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4321 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4381 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4441 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4501 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4561 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4621 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4681 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4741 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4801 tcggatggaa gccggtcttg tcgatcagga tgatctggac g
//
