LOCUS       sequence-107137-        6497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses beta actin with both a PAGFP and mRFP tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  AUTHORS   Kueh HY, Charras GT, Mitchison TJ, Brieher WM
  TITLE     Actin disassembly by cofilin, coronin, and Aip1 occurs in bursts and
            is inhibited by barbed-end cappers.
  JOURNAL   J Cell Biol. 2008 Jul 28;182(2):341-53. doi: 10.1083/jcb.200801027.
  PUBMED    18663144
REFERENCE   2  (bases 1 to 6497)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2008-07-28- 28"; volume: "182"; 
            issue: "2"; pages: "341-53"
COMMENT     Sequence Label: Actin-mRFP-PAGFP
FEATURES             Location/Qualifiers
     source          1..6497
                     /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             597..1271
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             597..1271
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(727..745)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     995..1014
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1189..1212
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             join(1293..1295,1296..1298,1299..2009)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1338..1359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1599..1618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1946..1967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3285..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3322..3341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3376..3395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3413..3868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3571)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3762..3781
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3895..3999
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4001..4358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4001..4192
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4024..4044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4209..4344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4393..5187
                     /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             4393..5187
                     /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(4447..4466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5057..5076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5375..5394)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5419..5466
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6284..6303
                     /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 gctagcatgg
      601 cctcctccga ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg
      661 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
      721 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
      781 cccctcagtt ccagtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
      841 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
      901 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
      961 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     1021 gctgggaggc ctccaccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     1081 agatgaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca
     1141 tggccaagaa gcccgtgcag ctgcccggcg cctacaagac cgacatcaag ctggacatca
     1201 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact
     1261 ccaccggcgc ctccggactc agatctcgag gcatggtgag caagggcgag gagctgttca
     1321 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1381 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1441 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcagc tacggcgtgc
     1501 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1561 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1621 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1681 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1741 acgtctatat catggccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1801 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1861 gcgacggccc cgtgctgctg cccgacaacc actacctgag ccaccagtcc gccctgagca
     1921 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1981 tcactctcgg catggacgag ctgtacaagg gcggcggcac tcgagctatg gatgatgata
     2041 tcgccgcgct cgtcgtcgac aacggctccg gcatgtgcaa ggccggcttc gcgggcgacg
     2101 atgccccccg ggccgtcttc ccctccatcg tggggcgccc caggcaccag ggcgtgatgg
     2161 tgggcatggg tcagaaggat tcctatgtgg gcgacgaggc ccagagcaag agaggcatcc
     2221 tcaccctgaa gtaccccatc gagcacggca tcgtcaccaa ctgggacgac atggagaaaa
     2281 tctggcacca caccttctac aatgagctgc gtgtggctcc cgaggagcac cccgtgctgc
     2341 tgaccgaggc ccccctgaac cccaaggcca accgcgagaa gatgacccag atcatgtttg
     2401 agaccttcaa caccccagcc atgtacgttg ctatccaggc tgtgctatcc ctgtacgcct
     2461 ctggccgtac cactggcatc gtgatggact ccggtgacgg ggtcacccac actgtgccca
     2521 tctacgaggg gtatgccctc ccccatgcca tcctgcgtct ggacctggct ggccgggacc
     2581 tgactgacta cctcatgaag atcctcaccg agcgcggcta cagcttcacc accacggccg
     2641 agcgggaaat cgtgcgtgac attaaggaga agctgtgcta cgtcgccctg gacttcgagc
     2701 aagagatggc cacggctgct tccagctcct ccctggagaa gagctacgag ctgcctgacg
     2761 gccaggtcat caccattggc aatgagcggt tccgctgccc tgaggcactc ttccagcctt
     2821 ccttcctggg catggagtcc tgtggcatcc acgaaactac cttcaactcc atcatgaagt
     2881 gtgacgtgga catccgcaaa gacctgtacg ccaacacagt gctgtctggc ggcaccacca
     2941 tgtaccctgg cattgccgac aggatgcaga aggagatcac tgccctggca cccagcacaa
     3001 tgaagatcaa gatcattgct cctcctgagc gcaagtactc cgtgtggatc ggcggctcca
     3061 tcctggcctc gctgtccacc ttccagcaga tgtggatcag caagcaggag tatgacgagt
     3121 ccggcccctc catcgtccac cgcaaatgct tctagggatc caccggatct agataactga
     3181 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3241 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3421 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3481 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3541 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3601 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3661 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3721 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3781 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3841 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3901 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3961 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4021 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4201 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4261 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4321 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4381 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4561 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4921 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5221 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5281 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5341 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5401 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5461 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5521 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5581 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5641 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5701 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtattaccgc catgcat
//
