LOCUS       sequence-410535-        6724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cry2(1-531)-mCherry-Profilin.S138A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6724)
  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 6724)
  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.S138A
FEATURES             Location/Qualifiers
     source          1..6724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             43..837
                     /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             43..837
                     /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(97..116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     707..726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1025..1044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1069..1116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1445..2033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1934..1953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2208..2511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2512..2715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2666..2686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2766..4259
                     /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      4377..4386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4383..5090
                     /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(4528..4546)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4796..4815
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5659..5780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5696..5715)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5750..5769
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5787..6242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5924..5945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6136..6155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6269..6373
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6375..6724,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6375..6566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6398..6418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6583..6718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
       61 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
      121 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
      181 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
      241 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
      301 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
      361 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
      421 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
      481 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
      541 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
      601 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
      661 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
      721 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
      781 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
      841 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
      901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
      961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     1021 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1081 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     1141 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     1201 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     1261 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     1321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1561 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2101 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     2161 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2221 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2281 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2341 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2401 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2461 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2521 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     2581 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     2641 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     2701 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
     2761 tcgagatgaa gatggacaaa aagactatag tttggtttag aagagaccta aggattgagg
     2821 ataatcctgc attagcagca gctgctcacg aaggatctgt ttttcctgtc ttcatttggt
     2881 gtcctgaaga agaaggacag ttttatcctg gaagagcttc aagatggtgg atgaaacaat
     2941 cacttgctca cttatctcaa tccttgaagg ctcttggatc tgacctcact ttaatcaaaa
     3001 cccacaacac gatttcagcg atcttggatt gtatccgcgt taccggtgct acaaaagtcg
     3061 tctttaacca cctctatgat cctgtttcgt tagttcggga ccataccgta aaggagaagc
     3121 tggtggaacg tgggatctct gtgcaaagct acaatggaga tctattgtat gaaccgtggg
     3181 agatatactg cgaaaagggc aaacctttta cgagtttcaa ttcttactgg aagaaatgct
     3241 tagatatgtc gattgaatcc gttatgcttc ctcctccttg gcggttgatg ccaataactg
     3301 cagcggctga agcgatttgg gcgtgttcga ttgaagaact agggctggag aatgaggccg
     3361 agaaaccgag caatgcgttg ttaactagag cttggtctcc aggatggagc aatgctgata
     3421 agttactaaa tgagttcatc gagaagcagt tgatagatta tgcaaagaac agcaagaaag
     3481 ttgttgggaa ttctacttca ctactttctc cgtatctcca tttcggggaa ataagcgtca
     3541 gacacgtttt ccagtgtgcc cggatgaaac aaattatatg ggcaagagat aagaacagtg
     3601 aaggagaaga aagtgcagat ctttttctta ggggaatcgg tttaagagag tattctcggt
     3661 atatatgttt caacttcccg tttactcacg agcaatcgtt gttgagtcat cttcggtttt
     3721 tcccttggga tgctgatgtt gataagttca aggcctggag acaaggcagg accggttatc
     3781 cgttggtgga tgccggaatg agagagcttt gggctaccgg atggatgcat aacagaataa
     3841 gagtgattgt ttcaagcttt gctgtgaagt ttcttctcct tccatggaaa tggggaatga
     3901 agtatttctg ggatacactt ttggatgctg atttggaatg tgacatcctt ggctggcagt
     3961 atatctctgg gagtatcccc gatggccacg agcttgatcg cttggacaat cccgcgttac
     4021 aaggcgccaa atatgaccca gaaggtgagt acataaggca atggcttccc gagcttgcga
     4081 gattgccaac tgaatggatc catcatccat gggacgctcc tttaaccgta ctcaaagctt
     4141 ctggtgtgga actcggaaca aactatgcga aacccattgt agacatcgac acagctcgtg
     4201 agctactagc taaagctatt tcaagaaccc gtgaagcaca gatcatgatc ggagcagcac
     4261 ctgatgagat tgtagcagat agcttcgagg ccttaggggc taataccatt aaagaacctg
     4321 gtctttgccc atctgtgtct tctaatgacc aacaagtagc ccgggatcca ccggtcgcca
     4381 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     4441 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     4501 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     4561 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     4621 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     4681 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     4741 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     4801 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     4861 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     4921 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     4981 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     5041 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggcggcagcg
     5101 gcggcggcat ggccgggtgg aacgcctaca tcgacagcct tatggcggac gggacctgtc
     5161 aggacgcggc catcgtaggc tacaaggact cgccctccgt ctgggccgcc gtccccggga
     5221 agaccttcgt tagcattacg ccagctgagg ttggtgtcct ggtaggcaaa gaccggtcaa
     5281 gttttttcgt caatgggctg acacttgggg gccagaaatg ttctgtgatc cgggactcac
     5341 tgctgcaaga cggggaattt acaatggatc ttcgtaccaa gagcaccgga ggagccccca
     5401 ccttcaatgt cactgtcacc atgactgcca agacgctagt cctgctgatg ggcaaagaag
     5461 gtgtccacgg tggtttgatc aacaagaaat gttatgaaat ggcctctcac ctgcggcgtg
     5521 cccagtacta ataatgagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     5581 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5641 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5701 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5761 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     5821 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5881 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5941 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     6001 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     6061 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     6121 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     6181 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     6241 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6301 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6361 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     6421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6481 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6541 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6601 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6661 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6721 ggct
//
