LOCUS       sequence-465607-        7564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  membrane localized EphrinA1 RTK fusion to Cry2olig - includes SAM 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7564)
  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 7564)
  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-SAM-Cry2olig
FEATURES             Location/Qualifiers
     source          1..7564
                     /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"
     CDS             537..557
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             1941..3434
                     /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      3453..3462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3459..4169
                     /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(3604..3622)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3872..3891
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4292..4413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4329..4348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4383..4402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4420..4875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4557..4578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4769..4788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4902..5006
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5008..5365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5008..5199
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5031..5051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5216..5351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5400..6194
                     /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             5400..6194
                     /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(5454..5473)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6064..6083
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6382..6401)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6426..6473
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6802..7390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7291..7310
                     /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 gctagcatgg
      541 gcagcagcaa aagcaaaccg aaaggcggca gcatggtgtc caggggcctg actggaggag
      601 agattgtagc cgtcatcttt gggctgctgc ttggtgcagc cttgctgctt gggattctcg
      661 ttttccggtc caggagagcc cagcggcaga ggcagcagag gcagcgtgac cgcgccaccg
      721 atgtggatcg agaggacaag ctgtggctga agccttatgt ggacctccag gcatacgagg
      781 accctgcaca gggagccttg gactttaccc gggagcttga tccagcgtgg ctgatggtgg
      841 acactgtcat aggagaagga gagtttgggg aagtgtatcg agggaccctg aggctcccca
      901 gccaggactg caagactgtg gccattaaga ccttaaaaga cacatcccca ggtggccagt
      961 ggtggaactt ccttcgagag gcaactatca tgggccagtt tagccacccg catattctgc
     1021 atctggaagg cgtcgtcaca aagcgaaagc cgatcatgat catcacagaa tttatggaga
     1081 atggagccct ggatgccttc ctgagggagc gggaggacca gctggtccct gggcagctag
     1141 tggccatgct gcagggcata gcatctggca tgaactacct cagtaatcac aattatgtcc
     1201 accgggacct ggctgccaga aacatcttgg tgaatcaaaa cctgtgctgc aaggtgtctg
     1261 actttggcct gactcgcctc ctggatgact ttgatggcac atacgaaacc cagggaggaa
     1321 agatccctat ccgttggaca gcccctgaag ccattgccca tcggatcttc accacagcca
     1381 gcgatgtgtg gagctttggg attgtgatgt gggaggtgct gagctttggg gacaagcctt
     1441 atggggagat gagcaatcag gaggttatga agagcattga ggatgggtac cggttgcccc
     1501 ctcctgtgga ctgccctgcc cctctgtatg agctcatgaa gaactgctgg gcatatgacc
     1561 gtgcccgccg gccacacttc cagaagcttc aggcacatct ggagcaactg cttgccaacc
     1621 cccactccct gcggaccatt gccaactttg accccagggt gactcttcgc ctgcccagcc
     1681 tgagtggctc agatgggatc ccgtatcgaa ccgtctctga gtggctcgag tccatacgca
     1741 tgaaacgcta catcctgcac ttccactcgg ctgggctgga caccatggag tgtgtgctgg
     1801 agctgaccgc tgaggacctg acgcagatgg gaatcacact gcccgggcac cagaagcgca
     1861 ttctttgcag tattcaggga ttcaaggact tgggcggcag cgactcgcta gcgctaccgg
     1921 actcagatct cgaggccacc atgaagatgg acaaaaagac tatagtttgg tttagaagag
     1981 acctaaggat tgaggataat cctgcattag cagcagctgc tcacgaagga tctgtttttc
     2041 ctgtcttcat ttggtgtcct gaagaagaag gacagtttta tcctggaaga gcttcaagat
     2101 ggtggatgaa acaatcactt gctcacttat ctcaatcctt gaaggctctt ggatctgacc
     2161 tcactttaat caaaacccac aacacgattt cagcgatctt ggattgtatc cgcgttaccg
     2221 gtgctacaaa agtcgtcttt aaccacctct atgatcctgt ttcgttagtt cgggaccata
     2281 ccgtaaagga gaagctggtg gaacgtggga tctctgtgca aagctacaat ggagatctat
     2341 tgtatgaacc gtgggagata tactgcgaaa agggcaaacc ttttacgagt ttcaattctt
     2401 actggaagaa atgcttagat atgtcgattg aatccgttat gcttcctcct ccttggcggt
     2461 tgatgccaat aactgcagcg gctgaagcga tttgggcgtg ttcgattgaa gaactagggc
     2521 tggagaatga ggccgagaaa ccgagcaatg cgttgttaac tagagcttgg tctccaggat
     2581 ggagcaatgc tgataagtta ctaaatgagt tcatcgagaa gcagttgata gattatgcaa
     2641 agaacagcaa gaaagttgtt gggaattcta cttcactact ttctccgtat ctccatttcg
     2701 gggaaataag cgtcagacac gttttccagt gtgcccggat gaaacaaatt atatgggcaa
     2761 gagataagaa cagtgaagga gaagaaagtg cagatctttt tcttagggga atcggtttaa
     2821 gagagtattc tcggtatata tgtttcaact tcccgtttac tcacgagcaa tcgttgttga
     2881 gtcatcttcg gtttttccct tgggatgctg atgttgataa gttcaaggcc tggagacaag
     2941 gcaggaccgg ttatccgttg gtggatgccg gaatgagaga gctttgggct accggatgga
     3001 tgcataacag aataagagtg attgtttcaa gctttgctgt gaagtttctt ctccttccat
     3061 ggaaatgggg aatgaagtat ttctgggata cacttttgga tgctgatttg gaatgtgaca
     3121 tccttggctg gcagtatatc tctgggagta tccccgatgg ccacgagctt gatcgcttgg
     3181 acaatcccgc gttacaaggc gccaaatatg acccagaagg tgagtacata aggcaatggc
     3241 ttcccgagct tgcgagattg ccaactgaat ggatccatca tccatgggac gctcctttaa
     3301 ccgtactcaa agcttctggt gtggaactcg gaacaaacta tgcgaaaccc attgtagaca
     3361 tcgacacagc tcgtgagcta ctagctaaag ctatttcaag aacccgtgga gcacagatca
     3421 tgatcggagc agcagcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg
     3481 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     3541 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3601 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     3661 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     3721 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3781 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     3841 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     3901 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     3961 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     4021 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     4081 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     4141 ccggcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     4201 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4261 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4381 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4441 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4501 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4561 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4621 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4681 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4741 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4801 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4861 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4921 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4981 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5041 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5101 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5161 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5221 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5281 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5341 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5401 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5461 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5521 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5581 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5641 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5701 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5761 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5821 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5881 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5941 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6001 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6061 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6121 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6181 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6241 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6301 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6361 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6421 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6481 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6541 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6601 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6661 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6721 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6781 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6841 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6901 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7261 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7321 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7381 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7441 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7501 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     7561 tccg
//
