LOCUS       sequence-348221-        7528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EphrinA1 RTK fusion to Cry2olig - includes SAM domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7528)
  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 7528)
  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: EphA1-SAM-Cry2olig
FEATURES             Location/Qualifiers
     source          1..7528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1369..2862
                     /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      2881..2890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2887..3597
                     /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(3032..3050)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3300..3319
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3720..3841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3757..3776)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3811..3830
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3848..4303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3985..4006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4197..4216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4330..4434
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4436..4793
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4436..4627
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4459..4479)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4644..4779
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4828..5622
                     /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             4828..5622
                     /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(4882..4901)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5492..5511
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5810..5829)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5854..5901
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6230..6818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6719..6738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6993..7296
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7297..7500
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7451..7471
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggtgtcca ggggcctgac tggaggagag attgtagccg tcatctttgg gctgctgctt
       61 ggtgcagcct tgctgcttgg gattctcgtt ttccggtcca ggagagccca gcggcagagg
      121 cagcagaggc agcgtgaccg cgccaccgat gtggatcgag aggacaagct gtggctgaag
      181 ccttatgtgg acctccaggc atacgaggac cctgcacagg gagccttgga ctttacccgg
      241 gagcttgatc cagcgtggct gatggtggac actgtcatag gagaaggaga gtttggggaa
      301 gtgtatcgag ggaccctgag gctccccagc caggactgca agactgtggc cattaagacc
      361 ttaaaagaca catccccagg tggccagtgg tggaacttcc ttcgagaggc aactatcatg
      421 ggccagttta gccacccgca tattctgcat ctggaaggcg tcgtcacaaa gcgaaagccg
      481 atcatgatca tcacagaatt tatggagaat ggagccctgg atgccttcct gagggagcgg
      541 gaggaccagc tggtccctgg gcagctagtg gccatgctgc agggcatagc atctggcatg
      601 aactacctca gtaatcacaa ttatgtccac cgggacctgg ctgccagaaa catcttggtg
      661 aatcaaaacc tgtgctgcaa ggtgtctgac tttggcctga ctcgcctcct ggatgacttt
      721 gatggcacat acgaaaccca gggaggaaag atccctatcc gttggacagc ccctgaagcc
      781 attgcccatc ggatcttcac cacagccagc gatgtgtgga gctttgggat tgtgatgtgg
      841 gaggtgctga gctttgggga caagccttat ggggagatga gcaatcagga ggttatgaag
      901 agcattgagg atgggtaccg gttgccccct cctgtggact gccctgcccc tctgtatgag
      961 ctcatgaaga actgctgggc atatgaccgt gcccgccggc cacacttcca gaagcttcag
     1021 gcacatctgg agcaactgct tgccaacccc cactccctgc ggaccattgc caactttgac
     1081 cccagggtga ctcttcgcct gcccagcctg agtggctcag atgggatccc gtatcgaacc
     1141 gtctctgagt ggctcgagtc catacgcatg aaacgctaca tcctgcactt ccactcggct
     1201 gggctggaca ccatggagtg tgtgctggag ctgaccgctg aggacctgac gcagatggga
     1261 atcacactgc ccgggcacca gaagcgcatt ctttgcagta ttcagggatt caaggacttg
     1321 ggcggcagcg actcgctagc gctaccggac tcagatctcg aggccaccat gaagatggac
     1381 aaaaagacta tagtttggtt tagaagagac ctaaggattg aggataatcc tgcattagca
     1441 gcagctgctc acgaaggatc tgtttttcct gtcttcattt ggtgtcctga agaagaagga
     1501 cagttttatc ctggaagagc ttcaagatgg tggatgaaac aatcacttgc tcacttatct
     1561 caatccttga aggctcttgg atctgacctc actttaatca aaacccacaa cacgatttca
     1621 gcgatcttgg attgtatccg cgttaccggt gctacaaaag tcgtctttaa ccacctctat
     1681 gatcctgttt cgttagttcg ggaccatacc gtaaaggaga agctggtgga acgtgggatc
     1741 tctgtgcaaa gctacaatgg agatctattg tatgaaccgt gggagatata ctgcgaaaag
     1801 ggcaaacctt ttacgagttt caattcttac tggaagaaat gcttagatat gtcgattgaa
     1861 tccgttatgc ttcctcctcc ttggcggttg atgccaataa ctgcagcggc tgaagcgatt
     1921 tgggcgtgtt cgattgaaga actagggctg gagaatgagg ccgagaaacc gagcaatgcg
     1981 ttgttaacta gagcttggtc tccaggatgg agcaatgctg ataagttact aaatgagttc
     2041 atcgagaagc agttgataga ttatgcaaag aacagcaaga aagttgttgg gaattctact
     2101 tcactacttt ctccgtatct ccatttcggg gaaataagcg tcagacacgt tttccagtgt
     2161 gcccggatga aacaaattat atgggcaaga gataagaaca gtgaaggaga agaaagtgca
     2221 gatctttttc ttaggggaat cggtttaaga gagtattctc ggtatatatg tttcaacttc
     2281 ccgtttactc acgagcaatc gttgttgagt catcttcggt ttttcccttg ggatgctgat
     2341 gttgataagt tcaaggcctg gagacaaggc aggaccggtt atccgttggt ggatgccgga
     2401 atgagagagc tttgggctac cggatggatg cataacagaa taagagtgat tgtttcaagc
     2461 tttgctgtga agtttcttct ccttccatgg aaatggggaa tgaagtattt ctgggataca
     2521 cttttggatg ctgatttgga atgtgacatc cttggctggc agtatatctc tgggagtatc
     2581 cccgatggcc acgagcttga tcgcttggac aatcccgcgt tacaaggcgc caaatatgac
     2641 ccagaaggtg agtacataag gcaatggctt cccgagcttg cgagattgcc aactgaatgg
     2701 atccatcatc catgggacgc tcctttaacc gtactcaaag cttctggtgt ggaactcgga
     2761 acaaactatg cgaaacccat tgtagacatc gacacagctc gtgagctact agctaaagct
     2821 atttcaagaa cccgtggagc acagatcatg atcggagcag cagcccggga tccaccggtc
     2881 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     2941 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     3001 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     3061 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     3121 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     3181 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     3241 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     3301 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     3361 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     3421 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     3481 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     3541 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaagc
     3601 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3661 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3721 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3781 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3841 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3901 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3961 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4021 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4081 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4141 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4201 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4261 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4381 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4441 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4561 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4621 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4681 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4741 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4801 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4861 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4921 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4981 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     5041 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5101 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5161 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5221 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5281 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5341 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5401 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5461 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5521 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5581 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5641 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5701 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5761 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     5821 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5881 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     5941 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     6001 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     6061 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6121 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6181 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6301 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6361 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6421 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6481 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6541 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6601 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6661 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6781 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6841 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6901 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     6961 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     7021 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     7081 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     7141 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     7201 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     7261 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     7321 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     7381 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     7441 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     7501 ggtttagtga accgtcagat ccgctagc
//
