LOCUS       sequence-218288-        5360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytosolic FRET, Excitation: 462 / 548, Emission: 492 /
            561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5360)
  AUTHORS   Davidson Lab
  TITLE     FRET Indicators - Live and Fixed Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5360)
  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: 
            "Unpublished"
COMMENT     Sequence Label: mTFP1-mKO-FRET-8
FEATURES             Location/Qualifiers
     source          1..5360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             578..1285
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     regulatory      1304..1313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1310..1966
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    2088..2209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2125..2144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2179..2198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2216..2671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2353..2374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2565..2584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2698..2802
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2804..3161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2804..2995
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2827..2847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3012..3147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3196..3990
                     /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             3196..3990
                     /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(3250..3269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3860..3879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4178..4197)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4222..4269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4598..5186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5087..5106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcaccatg gtgagcaagg gcgaggagac
      601 cacaatgggc gtaatcaagc ccgacatgaa gatcaagctg aagatggagg gcaacgtgaa
      661 tggccacgcc ttcgtgatcg agggcgaggg cgagggcaag ccctacgacg gcaccaacac
      721 catcaacctg gaggtgaagg agggagcccc cctgcccttc tcctacgaca ttctgaccac
      781 cgcgttcgcc tacggcaaca gggccttcac caagtacccc gacgacatcc ccaactactt
      841 caagcagtcc ttccccgagg gctactcttg ggagcgcacc atgaccttcg aggacaaggg
      901 catcgtgaag gtgaagtccg acatctccat ggaggaggac tccttcatct acgagataca
      961 cctcaagggc gagaacttcc cccccaacgg ccccgtgatg cagaagaaga ccaccggctg
     1021 ggacgcctcc accgagagga tgtacgtgcg cgacggcgtg ctgaagggcg acgtcaagca
     1081 caagctgctg ctggagggcg gcggccacca ccgcgttgac ttcaagacca tctacagggc
     1141 caagaaggcg gtgaagctgc ccgactatca ctttgtggac caccgcatcg agatcctgaa
     1201 ccacgacaag gactacaaca aggtgaccgt ttacgagagc gccgtggccc gcaactccac
     1261 cgacggcatg gacgagctgt acaagggggc ctcaggaccg gtcgccacca tggtgagtgt
     1321 gattaaacca gagatgaaga tgaggtacta catggacggc tccgtcaatg ggcatgagtt
     1381 cacaattgaa ggtgaaggca caggcagacc ttacgaggga catcaagaga tgacactacg
     1441 cgtcacaatg gccaagggcg ggccaatgcc tttcgcgttt gacttagtgt cacacgtgtt
     1501 ctgttacggc cacagacctt ttactaaata tccagaagag ataccagact atttcaaaca
     1561 agcatttcct gaaggcctgt catgggaaag gtcgttggag ttcgaagatg gtgggtccgc
     1621 ttcagtcagt gcgcatataa gccttagagg aaacaccttc taccacaaat ccaaatttac
     1681 tggggttaac tttcctgccg atggtcctat catgcaaaac caaagtgttg attgggagcc
     1741 atcaaccgag aaaattactg ccagcgacgg agttctgaag ggtgatgtta cgatgtacct
     1801 aaaacttgaa ggaggcggca atcacaaatg ccaattcaag actacttaca aggcggcaaa
     1861 aaagattctt aaaatgccag gaagccatta catcagccat cgcctcgtca ggaaaaccga
     1921 aggcaacatt actgagctgg tagaagatgc agtagctcat tcctaagcgg ccgcgactct
     1981 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2041 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2101 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2161 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2221 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2281 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2341 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2401 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2461 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2521 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2581 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2641 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2701 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2761 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2821 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2881 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3001 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3061 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3121 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3181 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3241 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3301 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3361 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3421 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3481 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3541 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3601 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3661 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3721 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3781 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3841 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3901 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3961 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4021 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4081 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4141 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4201 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4261 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4321 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4381 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4441 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4501 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4561 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4621 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4681 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4741 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4801 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4861 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4921 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4981 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5041 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5101 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5161 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5221 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5281 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     5341 agcccatata tggagttccg
//
