LOCUS       sequence-353406-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCerulean3 has S208F/V224L mutations that promote weak dimerization 
            with YPet to optimize oritentation for FRET. tag is SsrA (AANDENY)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  AUTHORS   Liu B, Stone OJ, Pablo M, Herron JC, Nogueira AT, Dagliyan O, Grimm 
            JB, Lavis LD, Elston TC, Hahn KM
  TITLE     Biosensors based on peptide exposure show single molecule 
            conformations in live cells.
  JOURNAL   Cell. 2021 Oct 28;184(22):5670-5685.e23. doi: 
            10.1016/j.cell.2021.09.026. Epub 2021 Oct 11.
  PUBMED    34637702
REFERENCE   2  (bases 1 to 6311)
  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: 
            "10.1016/j.cell.2021.09.026"; journalName: "Cell"; date: 
            "2021-10-28- 28"; volume: "184"; issue: "22"; pages: "5670-5685.e23"
COMMENT     Sequence Label: tagSrc(WT)-mCer3 S208F/V224L
FEATURES             Location/Qualifiers
     source          1..6311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        605..908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        909..1112
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1063..1083
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1149..1158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2799..2801,2802..3512)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIKA
                     NFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLFKDPNEKRDHMVLLE
                     FLTAAGITLGMDELYK"
     primer_bind     complement(2841..2862)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3102..3121)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3643..3764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3680..3699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3734..3753
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3771..4226)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3908..3929)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4120..4139
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4253..4357
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4359..4716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4359..4550
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4382..4402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4567..4702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4751..5545
                     /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             4751..5545
                     /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(4805..4824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5415..5434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5733..5752)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5777..5824
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(6153..6311,1..430)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
       61 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      481 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
      541 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      601 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      661 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      721 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      781 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      841 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      901 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      961 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1021 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     1081 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
     1141 ttaattaagc caccatgggc agcaacaaga gcaagcccaa ggacgccagc cagcggcgcc
     1201 gcagcctgga gccctcggaa aacgtgcacg gggcaggggg cgccttcccg gcctcacaga
     1261 caccgagcaa gcccgcctcc gccgacggcc accgcgggcc cagcgccgcc ttcgtgccgc
     1321 ccgcggccga gcccaagctc ttcggaggct tcaactcctc ggacaccgtc acctccccgc
     1381 agagggcggg gcctctggca ggtggggtga ccacctttgt ggccctctat gactatgagt
     1441 cacggacaga gactgacctg tccttcaaga aaggggagcg gctgcagatt gttaacaaca
     1501 cggagggaga ctggtggctg gcacactcgc tgagcacggg acagaccggt tacatcccca
     1561 gcaactatgt ggcgccctcc gactccatcc aggctgagga gtggtacttt ggcaagatca
     1621 ctagacggga atcagagcgg ctgctgctca acgccgagaa cccgagaggg accttcctcg
     1681 tgagggagag tgagaccaca aaaggtgcct actgcctctc tgtatccgac ttcgacaatg
     1741 ccaagggcct aaatgtgaaa cactacaaga tccgcaagct ggacagcggc ggtttctaca
     1801 tcacctcccg cacccagttc aacagcctgc agcagctcgt ggcttactac tccaaacatg
     1861 ctgatggcct gtgtcaccgc ctcactaccg tatgtcccac atccaagcct cagacccagg
     1921 gattggccaa ggatgcgtgg gagatccccc gggagtccct gcggctggag gtcaagctgg
     1981 gccagggttg cttcggagag gtgtggatgg ggacctggaa cggcaccacg agggttgcca
     2041 tcaaaactct gaagccaggc accatgtccc cagaggcctt cctgcaggag gcccaagtca
     2101 tgaagaaact taggcacgag aaactggtgc agctgtatgc tgtggtgtcg gaagaaccca
     2161 tttacattgt gacagagtac atgaacaagg ggagtctgct ggactttctc aagggggaaa
     2221 cgggcaaata tttgcggcta ccccagctgg tggacatgtc tgctcagatc gcttcaggca
     2281 tggcctatgt ggagcggatg aactatgtgc accgggacct tcgagccgcc aatatcctag
     2341 taggggagaa cctggtgtgc aaagtggccg actttgggtt ggcccggctc atagaagaca
     2401 acgaatacac agcccggcaa ggtgccaaat tccccatcaa gtggaccgcc cctgaagctg
     2461 ctctgtacgg caggttcacc atcaagtcgg atgtgtggtc ctttgggatt ctgctgaccg
     2521 agctcaccac taagggaaga gtgccctatc ctgggatggt gaaccgtgag gttctggacc
     2581 aggtggagcg gggctaccgg atgccttgtc cccccgagtg ccccgagtcc ctgcatgacc
     2641 ttatgtgcca gtgctggcgg aaggagcccg aggagcggcc caccttcgag tacctgcagg
     2701 ccttcctgga agactacttt acgtccactg agccacagta ccagcccggg gagaacgctg
     2761 caaatgatga aaactaccta ggcggaccag ggaagcttgt gagcaagggc gaggagctgt
     2821 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2881 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2941 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg tcctggggcg
     3001 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3061 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3121 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3181 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac gccatccacg
     3241 gcaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcggcctca
     3301 actgcaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3361 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctgt
     3421 tcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcttg accgccgccg
     3481 ggatcactct cggcatggac gagctgtaca aggaattcta agcggccgcg actctagatc
     3541 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3601 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3661 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3721 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3781 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3841 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3901 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3961 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4021 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4081 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4141 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4201 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4321 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4621 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4681 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4741 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4801 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4861 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4921 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4981 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5041 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5101 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5161 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5221 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5281 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5341 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5401 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5461 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5521 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5581 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5641 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5701 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5761 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5821 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5881 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5941 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6121 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6181 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6241 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6301 cttctagtgt a
//
