LOCUS       sequence-182373-        7578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses kinase-dead mutant of OptoEphB2 labeled with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7578)
  TITLE     optoEphB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7578)
  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: 
            "Unpublished"
COMMENT     Sequence Label: OptoEphB2KD-mCherry
FEATURES             Location/Qualifiers
     source          1..7578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    563..587
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             608..628
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     protein_bind    complement(1914..1938)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1955..3448
                     /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
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     regulatory      3467..3476
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3473..4183
                     /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(3618..3636)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3886..3905
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4306..4427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4343..4362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4397..4416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4434..4889)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4571..4592)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4783..4802
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4916..5020
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5022..5379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5022..5213
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5045..5065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5230..5365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5414..6208
                     /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             5414..6208
                     /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(5468..5487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6078..6097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6396..6415)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6440..6487
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6816..7404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7305..7324
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgaatc aaacaagttt gtacaaaaaa gcaggctccg cggccgcccc
      601 cttcaccatg ggaagcagca agagcaagcc aaagctttgt aacagacggg ggtttgagcg
      661 tgctgactcg gagtacacgg acaagctgca acactacacc agtggccaca tgaccccagg
      721 catgaagatc tacatcgatc ctttcaccta cgaggacccc aacgaggcag tgcgggagtt
      781 tgccaaggaa attgacatct cctgtgtcaa aattgagcag gtgatcggag caggggagtt
      841 tggcgaggtc tgcagtggcc acctgaagct gccaggcaag agagagatct ttgtggccat
      901 catgacgctc aagtcgggct acacggagaa gcagcgccgg gacttcctga gcgaagcctc
      961 catcatgggc cagttcgacc atcccaacgt catccacctg gagggtgtcg tgaccaagag
     1021 cacacctgtg atgatcatca ccgagttcat ggagaatggc tccctggact cctttctccg
     1081 gcaaaacgat gggcagttca cagtcatcca gctggtgggc atgcttcggg gcatcgcagc
     1141 tggcatgaag tacctggcag acatgaacta tgttcaccgt gacctggctg cccgcaacat
     1201 cctcgtcaac agcaacctgg tctgcaaggt gtcggacttt gggctctcac gctttctaga
     1261 ggacgatacc tcagacccca cctacaccag tgccctgggc ggaaagatcc ccatccgctg
     1321 gacagccccg gaagccatcc agtaccggaa gttcacctcg gccagtgatg tgtggagcta
     1381 cggcattgtc atgtgggagg tgatgtccta tggggagcgg ccctactggg acatgaccaa
     1441 ccaggatgta atcaatgcca ttgagcagga ctatcggctg ccaccgccca tggactgccc
     1501 gagcgccctg caccaactca tgctggactg ttggcagaag gaccgcaacc accggcccaa
     1561 gttcggccaa attgtcaaca cgctagacaa gatgatccgc aatcccaaca gcctcaaagc
     1621 catggcgccc ctctcctctg gcatcaacct gccgctgctg gaccgcacga tccccgacta
     1681 caccagcttt agcacggtgg acgagtggct ggaggccatc aagatggggc agtacaagga
     1741 gagcttcgcc aatgccggct tcacctcctt tgacgtcgtg tctcagatga tgatggagga
     1801 cattctccgg gttggggtca ctttggctgg ccaccagaaa aaaatcctga acagtatcca
     1861 ggtgatgcgg gcgcagatga accagattca gtctgtggag gtttggcgcg ccgacccagc
     1921 tttcttgtac aaagtggttc gattcgaggc caccatgaag atggacaaaa agactatagt
     1981 ttggtttaga agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga
     2041 aggatctgtt tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg
     2101 aagagcttca agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc
     2161 tcttggatct gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg
     2221 tatccgcgtt accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt
     2281 agttcgggac cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta
     2341 caatggagat ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac
     2401 gagtttcaat tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc
     2461 tcctccttgg cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat
     2521 tgaagaacta gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc
     2581 ttggtctcca ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt
     2641 gatagattat gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc
     2701 gtatctccat ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca
     2761 aattatatgg gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag
     2821 gggaatcggt ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga
     2881 gcaatcgttg ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa
     2941 ggcctggaga caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg
     3001 ggctaccgga tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt
     3061 tcttctcctt ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga
     3121 tttggaatgt gacatccttg gctggcagta tatctctggg agtatccccg atggccacga
     3181 gcttgatcgc ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta
     3241 cataaggcaa tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg
     3301 ggacgctcct ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa
     3361 acccattgta gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg
     3421 tggagcacag atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatggtgag
     3481 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     3541 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     3601 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     3661 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     3721 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     3781 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     3841 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     3901 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     3961 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     4021 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     4081 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     4141 gggccgccac tccaccggcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     4201 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4261 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4321 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4381 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     4441 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4501 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4561 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4621 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4681 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4741 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4801 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4861 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4981 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     5041 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5101 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5161 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5281 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5341 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     5401 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5461 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5521 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5581 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5641 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5701 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5761 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5821 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5881 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5941 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6001 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6061 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6121 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6181 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6241 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6301 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6361 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     6421 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     6481 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     6541 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     6601 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6661 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6721 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6781 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6841 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6901 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6961 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7021 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7081 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7141 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7201 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7261 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7321 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7381 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7441 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7501 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     7561 cccatatatg gagttccg
//
