LOCUS       sequence-410531-        6724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cry2(1-531)-mCherry-Profilin.S138E.
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.S138E
FEATURES             Location/Qualifiers
     source          1..6724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             80..874
                     /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             80..874
                     /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(134..153)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     744..763
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1062..1081)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1106..1153
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1482..2070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1971..1990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2245..2548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2549..2752
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2703..2723
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2803..4296
                     /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      4414..4423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4420..5127
                     /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(4565..4583)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4833..4852
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5696..5817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5733..5752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5787..5806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5824..6279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5961..5982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6173..6192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6306..6410
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6412..6724,1..45)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6412..6603
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6435..6455)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6620..6724,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
       61 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
      121 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
      181 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
      241 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
      301 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
      361 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
      421 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
      481 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
      541 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
      601 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
      661 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
      721 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
      781 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
      841 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
      901 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
      961 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     1021 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     1081 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     1141 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     1201 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     1261 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     1321 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1381 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1621 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2101 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2161 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     2221 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     2281 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     2341 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     2401 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     2461 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     2521 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     2581 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     2641 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     2701 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     2761 gaaccgtcag atccgctagc gctaccggac tcagatctcg agatgaagat ggacaaaaag
     2821 actatagttt ggtttagaag agacctaagg attgaggata atcctgcatt agcagcagct
     2881 gctcacgaag gatctgtttt tcctgtcttc atttggtgtc ctgaagaaga aggacagttt
     2941 tatcctggaa gagcttcaag atggtggatg aaacaatcac ttgctcactt atctcaatcc
     3001 ttgaaggctc ttggatctga cctcacttta atcaaaaccc acaacacgat ttcagcgatc
     3061 ttggattgta tccgcgttac cggtgctaca aaagtcgtct ttaaccacct ctatgatcct
     3121 gtttcgttag ttcgggacca taccgtaaag gagaagctgg tggaacgtgg gatctctgtg
     3181 caaagctaca atggagatct attgtatgaa ccgtgggaga tatactgcga aaagggcaaa
     3241 ccttttacga gtttcaattc ttactggaag aaatgcttag atatgtcgat tgaatccgtt
     3301 atgcttcctc ctccttggcg gttgatgcca ataactgcag cggctgaagc gatttgggcg
     3361 tgttcgattg aagaactagg gctggagaat gaggccgaga aaccgagcaa tgcgttgtta
     3421 actagagctt ggtctccagg atggagcaat gctgataagt tactaaatga gttcatcgag
     3481 aagcagttga tagattatgc aaagaacagc aagaaagttg ttgggaattc tacttcacta
     3541 ctttctccgt atctccattt cggggaaata agcgtcagac acgttttcca gtgtgcccgg
     3601 atgaaacaaa ttatatgggc aagagataag aacagtgaag gagaagaaag tgcagatctt
     3661 tttcttaggg gaatcggttt aagagagtat tctcggtata tatgtttcaa cttcccgttt
     3721 actcacgagc aatcgttgtt gagtcatctt cggtttttcc cttgggatgc tgatgttgat
     3781 aagttcaagg cctggagaca aggcaggacc ggttatccgt tggtggatgc cggaatgaga
     3841 gagctttggg ctaccggatg gatgcataac agaataagag tgattgtttc aagctttgct
     3901 gtgaagtttc ttctccttcc atggaaatgg ggaatgaagt atttctggga tacacttttg
     3961 gatgctgatt tggaatgtga catccttggc tggcagtata tctctgggag tatccccgat
     4021 ggccacgagc ttgatcgctt ggacaatccc gcgttacaag gcgccaaata tgacccagaa
     4081 ggtgagtaca taaggcaatg gcttcccgag cttgcgagat tgccaactga atggatccat
     4141 catccatggg acgctccttt aaccgtactc aaagcttctg gtgtggaact cggaacaaac
     4201 tatgcgaaac ccattgtaga catcgacaca gctcgtgagc tactagctaa agctatttca
     4261 agaacccgtg aagcacagat catgatcgga gcagcacctg atgagattgt agcagatagc
     4321 ttcgaggcct taggggctaa taccattaaa gaacctggtc tttgcccatc tgtgtcttct
     4381 aatgaccaac aagtagcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     4441 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     4501 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4561 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     4621 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     4681 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     4741 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     4801 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     4861 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     4921 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     4981 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     5041 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     5101 accggcggca tggacgagct gtacaagggc ggcagcggcg gcggcatggc cgggtggaac
     5161 gcctacatcg acagccttat ggcggacggg acctgtcagg acgcggccat cgtaggctac
     5221 aaggactcgc cctccgtctg ggccgccgtc cccgggaaga ccttcgttag cattacgcca
     5281 gctgaggttg gtgtcctggt aggcaaagac cggtcaagtt ttttcgtcaa tgggctgaca
     5341 cttgggggcc agaaatgttc tgtgatccgg gactcactgc tgcaagacgg ggaatttaca
     5401 atggatcttc gtaccaagag caccggagga gcccccacct tcaatgtcac tgtcaccatg
     5461 actgccaaga cgctagtcct gctgatgggc aaagaaggtg tccacggtgg tttgatcaac
     5521 aagaaatgtt atgaaatggc ctctcacctg cggcgtgaac agtactaata atgagcggcc
     5581 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5641 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5701 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5761 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     5821 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5881 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5941 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6001 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6061 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6121 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6181 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6241 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     6301 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6361 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     6421 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6481 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6541 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6601 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6661 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6721 attc
//
