LOCUS       sequence-410533-        6723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cry2(1-531)-mCherry-Profilin.WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6723)
  AUTHORS   Mann N, Hill J, Wang K, Hughes RM
  TITLE     OptoProfilin: A Single Component Biosensor of Applied Cellular 
            Stress.
  JOURNAL   Chembiochem. 2024 May 2;25(9):e202400007. doi: 
            10.1002/cbic.202400007. Epub 2024 Mar 28.
  PUBMED    38457348
REFERENCE   2  (bases 1 to 6723)
  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: 
            "Chembiochem."; date: "2024-05-2"; volume: "25(9):e202400007. doi"; 
            pages: " 10.1002/cbic.202400007. Epub 2024 Mar 28"
COMMENT     Sequence Label: OptoProfilin
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             137..931
                     /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             137..931
                     /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(191..210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     801..820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1119..1138)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1163..1210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1539..2127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2028..2047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2302..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2606..2809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2860..4353
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     regulatory      4471..4480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4477..5184
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4622..4640)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4890..4909
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5752..5873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5789..5808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5843..5862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5880..6335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6017..6038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6229..6248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6362..6466
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6468..6723,1..102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6468..6659
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6491..6511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6676..6723,1..88)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
       61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
      121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
      181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
      241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
      301 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
      361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
      421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
      481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
      541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
      601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
      661 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
      721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
      781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
      841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
      901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
      961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     1021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1081 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     1141 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     1201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     1261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     1321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     1381 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2221 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2821 ccgtcagatc cgctagcgct accggactca gatctcgaga tgaagatgga caaaaagact
     2881 atagtttggt ttagaagaga cctaaggatt gaggataatc ctgcattagc agcagctgct
     2941 cacgaaggat ctgtttttcc tgtcttcatt tggtgtcctg aagaagaagg acagttttat
     3001 cctggaagag cttcaagatg gtggatgaaa caatcacttg ctcacttatc tcaatccttg
     3061 aaggctcttg gatctgacct cactttaatc aaaacccaca acacgatttc agcgatcttg
     3121 gattgtatcc gcgttaccgg tgctacaaaa gtcgtcttta accacctcta tgatcctgtt
     3181 tcgttagttc gggaccatac cgtaaaggag aagctggtgg aacgtgggat ctctgtgcaa
     3241 agctacaatg gagatctatt gtatgaaccg tgggagatat actgcgaaaa gggcaaacct
     3301 tttacgagtt tcaattctta ctggaagaaa tgcttagata tgtcgattga atccgttatg
     3361 cttcctcctc cttggcggtt gatgccaata actgcagcgg ctgaagcgat ttgggcgtgt
     3421 tcgattgaag aactagggct ggagaatgag gccgagaaac cgagcaatgc gttgttaact
     3481 agagcttggt ctccaggatg gagcaatgct gataagttac taaatgagtt catcgagaag
     3541 cagttgatag attatgcaaa gaacagcaag aaagttgttg ggaattctac ttcactactt
     3601 tctccgtatc tccatttcgg ggaaataagc gtcagacacg ttttccagtg tgcccggatg
     3661 aaacaaatta tatgggcaag agataagaac agtgaaggag aagaaagtgc agatcttttt
     3721 cttaggggaa tcggtttaag agagtattct cggtatatat gtttcaactt cccgtttact
     3781 cacgagcaat cgttgttgag tcatcttcgg tttttccctt gggatgctga tgttgataag
     3841 ttcaaggcct ggagacaagg caggaccggt tatccgttgg tggatgccgg aatgagagag
     3901 ctttgggcta ccggatggat gcataacaga ataagagtga ttgtttcaag ctttgctgtg
     3961 aagtttcttc tccttccatg gaaatgggga atgaagtatt tctgggatac acttttggat
     4021 gctgatttgg aatgtgacat ccttggctgg cagtatatct ctgggagtat ccccgatggc
     4081 cacgagcttg atcgcttgga caatcccgcg ttacaaggcg ccaaatatga cccagaaggt
     4141 gagtacataa ggcaatggct tcccgagctt gcgagattgc caactgaatg gatccatcat
     4201 ccatgggacg ctcctttaac cgtactcaaa gcttctggtg tggaactcgg aacaaactat
     4261 gcgaaaccca ttgtagacat cgacacagct cgtgagctac tagctaaagc tatttcaaga
     4321 acccgtgaag cacagatcat gatcggagca gcacctgatg agattgtagc agatagcttc
     4381 gaggccttag gggctaatac cattaaagaa cctggtcttt gcccatctgt gtcttctaat
     4441 gaccaacaag tagcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggaggat
     4501 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     4561 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     4621 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     4681 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     4741 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     4801 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     4861 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     4921 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     4981 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     5041 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     5101 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     5161 ggcggcatgg acgagctgta caagggcggc agcggcggcg gcatggccgg gtggaacgcc
     5221 tacatcgaca gccttatggc ggacgggacc tgtcaggacg cggccatcgt aggctacaag
     5281 gactcgccct ccgtctgggc cgccgtcccc gggaagacct tcgttagcat tacgccagct
     5341 gaggttggtg tcctggtagg caaagaccgg tcaagttttt tcgtcaatgg gctgacactt
     5401 gggggccaga aatgttctgt gatccgggac tcactgctgc aagacgggga atttacaatg
     5461 gatcttcgta ccaagagcac cggaggagcc cccaccttca atgtcactgt caccatgact
     5521 gccaagacgc tagtcctgct gatgggcaaa gaaggtgtcc acggtggttt gatcaacaag
     5581 aaatgttatg aaatggcctc tcacctgcgg cgttcccagt actaatatga gcggccgcga
     5641 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     5701 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     5761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     5881 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5941 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6001 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6061 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6121 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6181 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6241 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6301 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     6361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     6481 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     6541 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     6601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     6661 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6721 ggc
//
