LOCUS       sequence-300369-        4968 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 4968)
  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 4968)
  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..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             848..1558
                     /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    1756..1877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1793..1812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1847..1866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1884..2339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2021..2042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2233..2252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2366..2470
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2472..2829
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2472..2663
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2495..2515)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2680..2815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2864..3658
                     /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             2864..3658
                     /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(2918..2937)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3528..3547
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3846..3865)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3890..3937
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4266..4854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4755..4774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaccaatac ccgactctat atatcataga cgacgaacga gctaacgaac gacaaacact
       61 aataccaccg tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      661 gctagcgcta ccggactcag atctcgagct caagcttatg ggcatggcgc gaccacgggg
      721 cgcggggccc tgcagccccg ggttggagcg ggctccgcgc cggagcgtcg gggagctgcg
      781 cctgctcttc gaagcgcgct gcgccgcagt cgccgccgca gcggccgcag ggtcggatcc
      841 accggtcatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt
      901 catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcagacc
      961 gtacgaggga gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc
     1021 ctttgacatt cttgccacgt cgttcatgta tggcagccgt acctttatca agtacccggc
     1081 cgacatccct gatttcttta aacagtcctt tactgagggt tttacttggg aaagagttac
     1141 gagatacgaa gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga
     1201 gctcgtctac aacgtcaagg tcagaggggt aaactttccc tccaatggtc ccgtgatgca
     1261 gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct
     1321 gagaggatac actgacatcg cactgaaagt tgatggtggt ggccatctgc actgcaactt
     1381 cgtgacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc
     1441 cgttgatcac cgcctggaaa ggatcgagga gagtgacaat gaaacctacg tagtgcaaag
     1501 agaagtggca gttgccaaat acagcaacct tggtggtggc atggacgagc tgtacaagag
     1561 atctcgagct caagcttcga attctggtaa aaagaagaaa aagaagtcaa agacaaagtg
     1621 tgtaattatg tagagcggcc gcgactctag atcataatca gccataccac atttgtagag
     1681 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1741 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1801 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1861 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     1921 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1981 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2041 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2101 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2161 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2221 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2281 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2341 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2401 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2461 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2581 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2641 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2701 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2761 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2821 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2881 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2941 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3001 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3061 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3121 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3181 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3241 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3301 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3361 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3421 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3481 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3541 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3601 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3661 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3721 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3781 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     3841 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3901 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3961 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4021 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4081 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4141 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4201 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4261 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4321 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4381 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4441 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4501 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4561 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4621 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4681 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4741 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4801 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4861 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4921 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
