LOCUS       sequence-165712-        7500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TG2 biosensor that allows for quantitative assessment of TG2 
            conformational changes in live cells using FRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7500)
  AUTHORS   Caron NS, Munsie LN, Keillor JW, Truant R
  TITLE     Using FLIM-FRET to measure conformational changes of 
            transglutaminase type 2 in live cells.
  JOURNAL   PLoS One. 2012;7(8):e44159. doi: 10.1371/journal.pone.0044159. Epub 
            2012 Aug 31.
  PUBMED    22952912
REFERENCE   2  (bases 1 to 7500)
  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: "PLoS One.";
            date: "2012"; volume: "7(8):e44159. doi"; pages: " 
            10.1371/journal.pone.0044159. Epub 2012 Aug 31"
COMMENT     Sequence Label: mCer-TG2-YFP
FEATURES             Location/Qualifiers
     source          1..7500
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFEIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3451..3453,3454..3456,3457..4170)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4288..4409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4325..4344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4379..4398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4416..4871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4553..4574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4765..4784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4898..5002
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5004..5361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5004..5195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5027..5047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5212..5347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5396..6190
                     /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             5396..6190
                     /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(5450..5469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6060..6079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6378..6397)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6422..6469
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6798..7386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7287..7306
                     /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 gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaacgcc atcagcgaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc gagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctggtggc ggagggatgg ccgaggagct ggtcttagag
     1381 aggtgtgatc tggagctgga gaccaatggc cgagaccacc acacggccga cctgtgccgg
     1441 gagaagctgg tggtgcgacg gggccagccc ttctggctga ccctgcactt tgagggccgc
     1501 aactacgagg ccagtgtaga cagtctcacc ttcagtgtcg tgaccggccc agcccctagc
     1561 caggaggccg ggaccaaggc ccgttttcca ctaagagatg ctgtggagga gggtgactgg
     1621 acagccaccg tggtggacca gcaagactgc accctctcgc tgcagctcac caccccggcc
     1681 aacgccccca tcggcctgta tcgcctcagc ctggaggcct ccactggcta ccagggatcc
     1741 agctttgtgc tgggccactt cattttgctc ttcaacgcct ggtgcccagc ggatgctgtg
     1801 tacctggact cggaagagga gcggcaggag tatgtcctca cccagcaggg ctttatctac
     1861 cagggctcgg ccaagttcat caagaacata ccttggaatt ttgggcagtt tgaagatggg
     1921 atcctagaca tctgcctgat ccttctagat gtcaacccca agttcctgaa gaacgccggc
     1981 cgtgactgct cccgccgcag cagccccgtc tacgtgggcc gggtggtgag tggcatggtc
     2041 aactgcaacg atgaccaggg tgtgctgctg ggacgctggg acaacaacta cggggacggc
     2101 gtcagcccca tgtcctggat cggcagcgtg gacatcctgc ggcgctggaa gaaccacggc
     2161 tgccagcgcg tcaagtatgg ccagtgctgg gtcttcgccg ccgtggcctg cacagtgctg
     2221 aggtgcctgg gcatccctac ccgcgtcgtg accaactaca actcggccca tgaccagaac
     2281 agcaaccttc tcatcgagta cttccgcaat gagtttgggg agatccaggg tgacaagagc
     2341 gagatgatct ggaacttcca ctgctgggtg gagtcgtgga tgaccaggcc ggacctgcag
     2401 ccggggtacg agggctggca ggccctggac ccaacgcccc aggagaagag cgaagggacg
     2461 tactgctgtg gcccagttcc agttcgtgcc atcaaggagg gcgacctgag caccaagtac
     2521 gatgcgccct ttgtctttgc ggaggtcaat gccgacgtgg tagactggat ccagcaggac
     2581 gatgggtctg tgcacaaatc catcaaccgt tccctgatcg ttgggctgaa gatcagcact
     2641 aagagcgtgg gccgagacga gcgggaggat atcacccaca cctacaaata cccagagggg
     2701 tcctcagagg agagggaggc cttcacaagg gcgaaccacc tgaacaaact ggccgagaag
     2761 gaggagacag ggatggccat gcggatccgt gtgggccaga gcatgaacat gggcagtgac
     2821 tttgacgtct ttgcccacat caccaacaac accgctgagg agtacgtctg ccgcctcctg
     2881 ctctgtgccc gcaccgtcag ctacaatggg atcttggggc ccgagtgtgg caccaagtac
     2941 ctgctcaacc tcaacctgga gcctttctct gagaagagcg ttcctctttg catcctctat
     3001 gagaaatacc gtgactgcct tacggagtcc aacctcatca aggtgcgggc cctcctcgtg
     3061 gagccagtta tcaacagcta cctgctggct gagagggacc tctacctgga gaatccagaa
     3121 atcaagatcc ggatccttgg ggagcccaag cagaaacgca agctggtggc tgaggtgtcc
     3181 ctgcagaacc cgctccctgt ggccctggaa ggctgcacct tcactgtgga gggggccggc
     3241 ctgactgagg agcagaagac ggtggagatc ccagaccccg tggaggcagg ggaggaagtt
     3301 aaggtgagaa tggacctgct gccgctccac atgggcctcc acaagctggt ggtgaacttc
     3361 gagagcgaca agctgaaggc tgtgaagggc ttccggaatg tcatcattgg ccccgccggt
     3421 ggcggagggg gtaccgcggg cccgggatcc atggtgagca agggcgagga gctgttcacc
     3481 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3541 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3601 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag
     3661 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3721 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3781 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3841 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3901 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3961 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4021 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     4081 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4141 actctcggca tggacgagct gtacaagtaa tctagataac tgatcataat cagccatacc
     4201 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4261 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4321 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4381 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     4441 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4501 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4561 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4621 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4681 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4741 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4801 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4861 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4981 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     5041 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5101 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5161 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5221 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5281 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5341 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5401 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5461 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5521 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5581 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5641 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5701 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5761 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5821 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5881 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5941 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6001 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6061 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6121 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6181 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6241 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6301 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6361 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6421 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6481 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6541 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6601 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6661 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6721 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6781 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6901 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     6961 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7021 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7081 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7141 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7201 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7261 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7321 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7381 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7441 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
