LOCUS       sequence-192825-        5443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Measure FRET between mTFP1 and ShadowG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5443)
  AUTHORS   Demeautis C, Sipieter F, Roul J, Chapuis C, Padilla-Parra S, Riquet 
            FB, Tramier M
  TITLE     Multiplexing PKA and ERK1
  JOURNAL   Sci Rep. 2017 Jan 20;7:41026. doi: 10.1038/srep41026.
  PUBMED    28106114
REFERENCE   2  (bases 1 to 5443)
  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: "Sci Rep."; 
            date: "2017-01-20"; volume: "7:41026. doi"; pages: " 
            10.1038/srep41026"
COMMENT     Sequence Label: pmTFP1-ShadowG
FEATURES             Location/Qualifiers
     source          1..5443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      721..730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             727..1434
                     /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"
     misc_feature    1435..1500
                     /label=MCS
                     /note="multiple cloning site"
     CDS             join(1504..1506,1507..1509,1510..2223)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1549..1570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1810..1829)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2157..2178
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2345..2466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2382..2401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2436..2455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2473..2928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2610..2631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2822..2841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2955..3059
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3061..3418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3061..3252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3084..3104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3269..3404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3453..4247
                     /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             3453..4247
                     /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(3507..3526)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4117..4136
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4435..4454)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4479..4526
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4855..5443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5344..5363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      721 gccaccatgg tgagcaaggg cgaggagacc acaatgggcg taatcaagcc cgacatgaag
      781 atcaagctga agatggaggg caacgtgaat ggccacgcct tcgtgatcga gggcgagggc
      841 gagggcaagc cctacgacgg caccaacacc atcaacctgg aggtgaagga gggagccccc
      901 ctgcccttct cctacgacat tctgaccacc gcgttcgcct acggcaacag ggccttcacc
      961 aagtaccccg acgacatccc caactacttc aagcagtcct tccccgaggg ctactcttgg
     1021 gagcgcacca tgaccttcga ggacaagggc atcgtgaagg tgaagtccga catctccatg
     1081 gaggaggact ccttcatcta cgagatacac ctcaagggcg agaacttccc ccccaacggc
     1141 cccgtgatgc agaagaagac caccggctgg gacgcctcca ccgagaggat gtacgtgcgc
     1201 gacggcgtgc tgaagggcga cgtcaagcac aagctgctgc tggagggcgg cggccaccac
     1261 cgcgttgact tcaagaccat ctacagggcc aagaaggcgg tgaagctgcc cgactatcac
     1321 tttgtggacc accgcatcga gatcctgaac cacgacaagg actacaacaa ggtgaccgtt
     1381 tacgagagcg ccgtggcccg caattccacc gacggcatgg acgagctgta caagtccgga
     1441 ctcagatctc gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc
     1501 ggaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1561 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1621 tacggcaagc tgaccctgaa gctcatctgc accaccggca agctgcccgt gccctggccc
     1681 accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cgcgctatcc agaccacatg
     1741 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1801 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1861 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1921 cacaagctcg agtacaactg gaacagccac aacgtctata tcatggccga caagcagaag
     1981 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2041 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2101 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     2161 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2221 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2281 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2341 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2401 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2461 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2521 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2581 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2641 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2701 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2761 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2821 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2881 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     2941 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3001 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3061 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3421 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3481 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3541 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3601 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     3661 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3721 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3781 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3841 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3901 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3961 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4021 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4081 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4141 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4201 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4261 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4321 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4381 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4441 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4501 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4561 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4621 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4681 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     4741 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4801 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4861 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4921 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4981 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5041 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5101 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5161 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5221 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5281 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5341 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5401 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaa
//
