LOCUS       sequence-348223-        7129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EphrinA1 RTK fusion to Cry2olig - RTK domain only.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7129)
  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 7129)
  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-RTK-Cry2olig
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             970..2463
                     /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      2482..2491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2488..3198
                     /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(2633..2651)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2901..2920
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3321..3442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3358..3377)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3412..3431
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3449..3904)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3586..3607)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3798..3817
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3931..4035
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4037..4394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4037..4228
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4060..4080)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4245..4380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4429..5223
                     /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             4429..5223
                     /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(4483..4502)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5093..5112
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5411..5430)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5455..5502
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6320..6339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6594..6897
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6898..7101
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7052..7072
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggaccctg cacagggagc cttggacttt acccgggagc ttgatccagc gtggctgatg
       61 gtggacactg tcataggaga aggagagttt ggggaagtgt atcgagggac cctgaggctc
      121 cccagccagg actgcaagac tgtggccatt aagaccttaa aagacacatc cccaggtggc
      181 cagtggtgga acttccttcg agaggcaact atcatgggcc agtttagcca cccgcatatt
      241 ctgcatctgg aaggcgtcgt cacaaagcga aagccgatca tgatcatcac agaatttatg
      301 gagaatggag ccctggatgc cttcctgagg gagcgggagg accagctggt ccctgggcag
      361 ctagtggcca tgctgcaggg catagcatct ggcatgaact acctcagtaa tcacaattat
      421 gtccaccggg acctggctgc cagaaacatc ttggtgaatc aaaacctgtg ctgcaaggtg
      481 tctgactttg gcctgactcg cctcctggat gactttgatg gcacatacga aacccaggga
      541 ggaaagatcc ctatccgttg gacagcccct gaagccattg cccatcggat cttcaccaca
      601 gccagcgatg tgtggagctt tgggattgtg atgtgggagg tgctgagctt tggggacaag
      661 ccttatgggg agatgagcaa tcaggaggtt atgaagagca ttgaggatgg gtaccggttg
      721 ccccctcctg tggactgccc tgcccctctg tatgagctca tgaagaactg ctgggcatat
      781 gaccgtgccc gccggccaca cttccagaag cttcaggcac atctggagca actgcttgcc
      841 aacccccact ccctgcggac cattgccaac tttgacccca gggtgactct tcgcctgccc
      901 agcctgagtg gctcagatgg gggcggcagc gactcgctag cgctaccgga ctcagatctc
      961 gaggccacca tgaagatgga caaaaagact atagtttggt ttagaagaga cctaaggatt
     1021 gaggataatc ctgcattagc agcagctgct cacgaaggat ctgtttttcc tgtcttcatt
     1081 tggtgtcctg aagaagaagg acagttttat cctggaagag cttcaagatg gtggatgaaa
     1141 caatcacttg ctcacttatc tcaatccttg aaggctcttg gatctgacct cactttaatc
     1201 aaaacccaca acacgatttc agcgatcttg gattgtatcc gcgttaccgg tgctacaaaa
     1261 gtcgtcttta accacctcta tgatcctgtt tcgttagttc gggaccatac cgtaaaggag
     1321 aagctggtgg aacgtgggat ctctgtgcaa agctacaatg gagatctatt gtatgaaccg
     1381 tgggagatat actgcgaaaa gggcaaacct tttacgagtt tcaattctta ctggaagaaa
     1441 tgcttagata tgtcgattga atccgttatg cttcctcctc cttggcggtt gatgccaata
     1501 actgcagcgg ctgaagcgat ttgggcgtgt tcgattgaag aactagggct ggagaatgag
     1561 gccgagaaac cgagcaatgc gttgttaact agagcttggt ctccaggatg gagcaatgct
     1621 gataagttac taaatgagtt catcgagaag cagttgatag attatgcaaa gaacagcaag
     1681 aaagttgttg ggaattctac ttcactactt tctccgtatc tccatttcgg ggaaataagc
     1741 gtcagacacg ttttccagtg tgcccggatg aaacaaatta tatgggcaag agataagaac
     1801 agtgaaggag aagaaagtgc agatcttttt cttaggggaa tcggtttaag agagtattct
     1861 cggtatatat gtttcaactt cccgtttact cacgagcaat cgttgttgag tcatcttcgg
     1921 tttttccctt gggatgctga tgttgataag ttcaaggcct ggagacaagg caggaccggt
     1981 tatccgttgg tggatgccgg aatgagagag ctttgggcta ccggatggat gcataacaga
     2041 ataagagtga ttgtttcaag ctttgctgtg aagtttcttc tccttccatg gaaatgggga
     2101 atgaagtatt tctgggatac acttttggat gctgatttgg aatgtgacat ccttggctgg
     2161 cagtatatct ctgggagtat ccccgatggc cacgagcttg atcgcttgga caatcccgcg
     2221 ttacaaggcg ccaaatatga cccagaaggt gagtacataa ggcaatggct tcccgagctt
     2281 gcgagattgc caactgaatg gatccatcat ccatgggacg ctcctttaac cgtactcaaa
     2341 gcttctggtg tggaactcgg aacaaactat gcgaaaccca ttgtagacat cgacacagct
     2401 cgtgagctac tagctaaagc tatttcaaga acccgtggag cacagatcat gatcggagca
     2461 gcagcccggg atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
     2521 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2581 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2641 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2701 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2761 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2821 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2881 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2941 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     3001 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     3061 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     3121 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     3181 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     3241 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3301 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3361 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3421 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3481 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3541 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3601 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3661 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3721 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3781 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3841 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3901 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3961 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4021 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4141 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4201 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4261 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4321 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4381 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4441 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4501 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4561 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4621 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4681 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4741 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4801 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4861 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4921 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4981 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5041 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5101 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5161 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5221 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5281 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5341 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5401 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5461 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5521 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5581 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5641 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6481 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     6541 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6601 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6661 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6721 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6781 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6841 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     6901 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     6961 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     7021 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     7081 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagc
//
