LOCUS       sequence-161459-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET acceptor for Cdc42 donor, flanked with the far-red FP mMaroon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  AUTHORS   Laviv T, Kim BB, Chu J, Lam AJ, Lin MZ, Yasuda R
  TITLE     Simultaneous dual-color fluorescence lifetime imaging with novel 
            red-shifted fluorescent proteins.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046.
  PUBMED    27798609
REFERENCE   2  (bases 1 to 5608)
  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: "Nat 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046."
COMMENT     Sequence Label: mMaroon-PAK-mMaroon
FEATURES             Location/Qualifiers
     source          1..5608
                     /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             613..1344
                     /codon_start=1
                     /product="bright, rapidly maturing, monomeric far-red 
                     fluorescent protein (Bajar et al., 2016)"
                     /label=mMaroon1
                     /translation="MVSKGEELIKENMHTKLYLTGTVNNHYFECTAEGEGKPYEGTQTN
                     RIKVVRGGPLPFAFDILAPCFMYGSKTFINHPPDIPDYFKQSFPEGFTWERTTVYEDGG
                     TLTATQDTSLQDGCLIYNVQVRGENFPSNGPVMQKKTLGWEASTETLYPADGSLEGRLD
                     WALKLVGGGHLHCRLETTYRSKKPAKNLKMPGVYFIDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             1540..2274
                     /codon_start=1
                     /product="bright, rapidly maturing, monomeric far-red 
                     fluorescent protein (Bajar et al., 2016)"
                     /label=mMaroon1
                     /translation="MVSKGEELIKENMHTKLYLTGTVNNHYFECTAEGEGKPYEGTQTN
                     RIKVVRGGPLPFAFDILAPCFMYGSKTFINHPPDIPDYFKQSFPEGFTWERTTVYEDGG
                     TLTATQDTSLQDGCLIYNVQVRGENFPSNGPVMQKKTLGWEASTETLYPADGSLEGRLD
                     WALKLVGGGHLHCRLETTYRSKKPAKNLKMPGVYFIDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2396..2517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2433..2452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2487..2506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2524..2979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2661..2682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2873..2892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3006..3110
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3112..3469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3112..3303
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3135..3155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3320..3455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3504..4298
                     /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             3504..4298
                     /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(3558..3577)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4168..4187
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4486..4505)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4530..4577
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4906..5494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5395..5414
                     /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 gagctgatca aggagaacat gcacaccaag
      661 ctgtacctga caggcaccgt gaacaaccac tacttcgaat gcaccgccga aggggagggc
      721 aagccctacg agggcaccca gaccaacagg atcaaggtgg tgcgcggagg ccccctgccg
      781 ttcgcattcg acatcctggc cccctgcttt atgtacggga gcaagacctt cattaaccat
      841 ccacctgaca ttcccgatta ttttaagcag tccttccctg agggcttcac atgggagaga
      901 accaccgttt acgaagacgg gggcacactc actgctacac aggacaccag cctccaggac
      961 ggctgcctga tctacaacgt ccaagtcaga ggggagaact tcccatccaa cggccctgtg
     1021 atgcagaaga aaacactcgg ctgggaggcc agtaccgaga ccctgtaccc cgctgacggc
     1081 agcctggaag gcagactgga ctgggccctg aagctcgtgg gcgggggcca cctgcactgc
     1141 agactggaaa ccacatacag atccaagaaa cccgcaaaga acctcaagat gcccggcgtc
     1201 tattttattg acaggagact ggaaagaatc aaggaggccg acaatgagac ctacgtcgag
     1261 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt
     1321 aatggcatgg acgagctgta caagtccgga ctcagatctc gaggaatgaa caaaaagaaa
     1381 gagaaagagc ggccagagat ttctctccct gcagattttg aacacacaat tcatgtcggt
     1441 gctgatgctg tcacagggga gtttacggga ataccagagc agtgggcccg cttgcttcag
     1501 acatcaaata tcactaagtt ggagcagaag ggatccggaa tggtgagcaa gggcgaggag
     1561 ctgatcaagg agaacatgca caccaagctg tacctgacag gcaccgtgaa caaccactac
     1621 ttcgaatgca ccgccgaagg ggagggcaag ccctacgagg gcacccagac caacaggatc
     1681 aaggtggtgc gcggaggccc cctgccgttc gcattcgaca tcctggcccc ctgctttatg
     1741 tacgggagca agaccttcat taaccatcca cctgacattc ccgattattt taagcagtcc
     1801 ttccctgagg gcttcacatg ggagagaacc accgtttacg aagacggggg cacactcact
     1861 gctacacagg acaccagcct ccaggacggc tgcctgatct acaacgtcca agtcagaggg
     1921 gagaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccagt
     1981 accgagaccc tgtaccccgc tgacggcagc ctggaaggca gactggactg ggccctgaag
     2041 ctcgtgggcg ggggccacct gcactgcaga ctggaaacca catacagatc caagaaaccc
     2101 gcaaagaacc tcaagatgcc cggcgtctat tttattgaca ggagactgga aagaatcaag
     2161 gaggccgaca atgagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac
     2221 ctccctagca aactggggca caaacttaat ggcatggacg agctgtacaa gtaagcggcc
     2281 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2341 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2401 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2461 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     2521 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2581 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2641 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2701 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2761 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2821 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2881 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2941 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3001 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3061 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3121 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3481 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3541 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3601 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3661 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3721 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3781 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3841 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3901 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3961 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4021 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4081 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4141 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4201 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4261 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4321 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4381 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4441 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4501 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4561 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4621 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4681 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4741 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4801 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5041 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5521 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5581 tgtggataac cgtattaccg ccatgcat
//
