LOCUS       sequence-348263-        7165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  membrane localized EphrinA1 RTK fusion to Cry2olig - RTK domain 
            only.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7165)
  AUTHORS   Wurz AI, Zheng KS, Hughes RM
  TITLE     Optogenetic Regulation of EphA1 RTK Activation and Signaling.
  JOURNAL   bioRxiv [Preprint]. 2024 Feb 7:2024.02.06.579139. doi: 
            10.1101/2024.02.06.579139.
  PUBMED    38370612
REFERENCE   2  (bases 1 to 7165)
  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: "bioRxiv 
            [Preprint]. 2024 Feb 7:2024.02.06.579139. doi: 
            10.1101/2024.02.06.579139."
COMMENT     Sequence Label: pm-EphA1-RTK-Cry2olig
FEATURES             Location/Qualifiers
     source          1..7165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..21
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             1006..2499
                     /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      2518..2527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2524..3234
                     /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(2669..2687)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2937..2956
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3357..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3394..3413)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3448..3467
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3485..3940)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3622..3643)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3834..3853
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3967..4071
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4073..4430
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4073..4264
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4096..4116)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4281..4416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4465..5259
                     /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             4465..5259
                     /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(4519..4538)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5129..5148
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5447..5466)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5491..5538
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5867..6455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6356..6375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6630..6933
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6934..7137
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7088..7108
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atgggcagca gcaaaagcaa accgaaaggc ggcagcatgg accctgcaca gggagccttg
       61 gactttaccc gggagcttga tccagcgtgg ctgatggtgg acactgtcat aggagaagga
      121 gagtttgggg aagtgtatcg agggaccctg aggctcccca gccaggactg caagactgtg
      181 gccattaaga ccttaaaaga cacatcccca ggtggccagt ggtggaactt ccttcgagag
      241 gcaactatca tgggccagtt tagccacccg catattctgc atctggaagg cgtcgtcaca
      301 aagcgaaagc cgatcatgat catcacagaa tttatggaga atggagccct ggatgccttc
      361 ctgagggagc gggaggacca gctggtccct gggcagctag tggccatgct gcagggcata
      421 gcatctggca tgaactacct cagtaatcac aattatgtcc accgggacct ggctgccaga
      481 aacatcttgg tgaatcaaaa cctgtgctgc aaggtgtctg actttggcct gactcgcctc
      541 ctggatgact ttgatggcac atacgaaacc cagggaggaa agatccctat ccgttggaca
      601 gcccctgaag ccattgccca tcggatcttc accacagcca gcgatgtgtg gagctttggg
      661 attgtgatgt gggaggtgct gagctttggg gacaagcctt atggggagat gagcaatcag
      721 gaggttatga agagcattga ggatgggtac cggttgcccc ctcctgtgga ctgccctgcc
      781 cctctgtatg agctcatgaa gaactgctgg gcatatgacc gtgcccgccg gccacacttc
      841 cagaagcttc aggcacatct ggagcaactg cttgccaacc cccactccct gcggaccatt
      901 gccaactttg accccagggt gactcttcgc ctgcccagcc tgagtggctc agatgggggc
      961 ggcagcgact cgctagcgct accggactca gatctcgagg ccaccatgaa gatggacaaa
     1021 aagactatag tttggtttag aagagaccta aggattgagg ataatcctgc attagcagca
     1081 gctgctcacg aaggatctgt ttttcctgtc ttcatttggt gtcctgaaga agaaggacag
     1141 ttttatcctg gaagagcttc aagatggtgg atgaaacaat cacttgctca cttatctcaa
     1201 tccttgaagg ctcttggatc tgacctcact ttaatcaaaa cccacaacac gatttcagcg
     1261 atcttggatt gtatccgcgt taccggtgct acaaaagtcg tctttaacca cctctatgat
     1321 cctgtttcgt tagttcggga ccataccgta aaggagaagc tggtggaacg tgggatctct
     1381 gtgcaaagct acaatggaga tctattgtat gaaccgtggg agatatactg cgaaaagggc
     1441 aaacctttta cgagtttcaa ttcttactgg aagaaatgct tagatatgtc gattgaatcc
     1501 gttatgcttc ctcctccttg gcggttgatg ccaataactg cagcggctga agcgatttgg
     1561 gcgtgttcga ttgaagaact agggctggag aatgaggccg agaaaccgag caatgcgttg
     1621 ttaactagag cttggtctcc aggatggagc aatgctgata agttactaaa tgagttcatc
     1681 gagaagcagt tgatagatta tgcaaagaac agcaagaaag ttgttgggaa ttctacttca
     1741 ctactttctc cgtatctcca tttcggggaa ataagcgtca gacacgtttt ccagtgtgcc
     1801 cggatgaaac aaattatatg ggcaagagat aagaacagtg aaggagaaga aagtgcagat
     1861 ctttttctta ggggaatcgg tttaagagag tattctcggt atatatgttt caacttcccg
     1921 tttactcacg agcaatcgtt gttgagtcat cttcggtttt tcccttggga tgctgatgtt
     1981 gataagttca aggcctggag acaaggcagg accggttatc cgttggtgga tgccggaatg
     2041 agagagcttt gggctaccgg atggatgcat aacagaataa gagtgattgt ttcaagcttt
     2101 gctgtgaagt ttcttctcct tccatggaaa tggggaatga agtatttctg ggatacactt
     2161 ttggatgctg atttggaatg tgacatcctt ggctggcagt atatctctgg gagtatcccc
     2221 gatggccacg agcttgatcg cttggacaat cccgcgttac aaggcgccaa atatgaccca
     2281 gaaggtgagt acataaggca atggcttccc gagcttgcga gattgccaac tgaatggatc
     2341 catcatccat gggacgctcc tttaaccgta ctcaaagctt ctggtgtgga actcggaaca
     2401 aactatgcga aacccattgt agacatcgac acagctcgtg agctactagc taaagctatt
     2461 tcaagaaccc gtggagcaca gatcatgatc ggagcagcag cccgggatcc accggtcgcc
     2521 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2581 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2641 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2701 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2761 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2821 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2881 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2941 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     3001 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     3061 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     3121 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3181 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaagcggc
     3241 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3301 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3361 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3421 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3481 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3541 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3601 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3661 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3721 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3781 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3841 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3901 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3961 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4021 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4081 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4201 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4261 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4321 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4381 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4441 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4501 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4561 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4621 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4681 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4741 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4801 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4861 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4921 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4981 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5041 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5101 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5161 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5221 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5281 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5341 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5401 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5461 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5521 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5581 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5641 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5701 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5761 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5821 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5881 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5941 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6001 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6061 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6481 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6541 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     6601 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     6661 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     6721 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6781 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6841 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6901 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     6961 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     7021 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     7081 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     7141 ttagtgaacc gtcagatccg ctagc
//
