LOCUS       sequence-310132-        4898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monitor changes in the density of membrane-proximal F-actin (MPA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4898)
  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 4898)
  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-mRuby3
FEATURES             Location/Qualifiers
     source          1..4898
                     /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             778..1488
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFTEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     polyA_signal    1686..1807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1723..1742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1777..1796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1814..2269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1951..1972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2163..2182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2296..2400
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2402..2759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2402..2593
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2425..2445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2610..2745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2794..3588
                     /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             2794..3588
                     /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(2848..2867)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3458..3477
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3776..3795)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3820..3867
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4196..4784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4685..4704
                     /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 ggcatggcgc gaccacgggg cgcggggccc
      661 tgcagccccg ggttggagcg ggctccgcgc cggagcgtcg gggagctgcg cctgctcttc
      721 gaagcgcgct gcgccgcagt cgccgccgca gcggccgcag ggtcggatcc accggtcatg
      781 gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt
      841 tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcagacc gtacgaggga
      901 gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt
      961 cttgccacgt cgttcatgta tggcagccgt acctttatca agtacccggc cgacatccct
     1021 gatttcttta aacagtcctt tactgagggt tttacttggg aaagagttac gagatacgaa
     1081 gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga gctcgtctac
     1141 aacgtcaagg tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc
     1201 aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct gagaggatac
     1261 actgacatcg cactgaaagt tgatggtggt ggccatctgc actgcaactt cgtgacaact
     1321 tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc cgttgatcac
     1381 cgcctggaaa ggatcgagga gagtgacaat gaaacctacg tagtgcaaag agaagtggca
     1441 gttgccaaat acagcaacct tggtggtggc atggacgagc tgtacaagag atctcgagct
     1501 caagcttcga attctggtaa aaagaagaaa aagaagtcaa agacaaagtg tgtaattatg
     1561 tagagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     1621 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1681 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1741 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1801 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     1861 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1921 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1981 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2041 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2101 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2161 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2221 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2401 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     2761 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2821 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2881 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2941 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3001 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3061 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3121 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3181 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3241 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3301 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3361 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3421 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3481 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3541 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3601 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3661 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3721 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     3781 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     3841 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     3901 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     3961 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4021 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4081 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4141 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4201 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4261 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4321 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4381 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4441 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4501 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4561 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4621 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4681 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4741 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4801 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4861 cccctgattc tgtggataac cgtattaccg ccatgcat
//
