LOCUS       sequence-367686-        7603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optically controlled eDrTrkA expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7603)
  AUTHORS   Leopold AV, Thankachan S, Yang C, Gerashchenko D, Verkhusha VV
  TITLE     A general approach for engineering RTKs optically controlled with 
            far-red light.
  JOURNAL   Nat Methods. 2022 Jul;19(7):871-880. doi: 
            10.1038/s41592-022-01517-z. Epub 2022 Jun 9.
  PUBMED    35681062
REFERENCE   2  (bases 1 to 7603)
  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"; doi: 
            "10.1038/s41592-022-01517-z"; journalName: "Nat Methods"; date: 
            "2022-07"; volume: "19"; issue: "7"; pages: "871-880"
COMMENT     Sequence Label: eDrTrkA
FEATURES             Location/Qualifiers
     source          1..7603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     597..656
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             660..1361
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(799..817)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1067..1086
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4391..4512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4428..4447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4482..4501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4519..4974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4656..4677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4868..4887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5001..5105
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5107..5464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5107..5298
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5130..5150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5315..5450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5499..6293
                     /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             5499..6293
                     /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(5553..5572)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6163..6182
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6481..6500)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6525..6572
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6901..7489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7390..7409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca
      661 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      721 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      781 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      841 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      901 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      961 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1021 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1081 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1141 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1201 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1261 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1321 agggccgcca ctccaccggc ggcatggacg agctgtacaa ggaattcagt gctggtagtg
     1381 ctggtagtgc tggcaccggt atgagccggg acccgttgcc cttttttcca ccgctttacc
     1441 ttggtggccc ggaaattacc accgagaact gcgagcgcga gccgattcat attcccggca
     1501 gcatccagcc gcacggcgcc ctgctcactg ccgacgggca cagcggcgag gtgctccaga
     1561 tgagcctcaa cgcggccact tttctgggac aggaacccac agtgctgcgc ggacagaccc
     1621 tcgccgcact gctgcccgag cagtggcccg cgctgcaagc ggccctgccc cccggctgcc
     1681 ccgacgccct gcaataccgc gcaacgctgg actggcctgc cgccgggcac ctttcgctga
     1741 cggtgcaccg ggtcggcgag ttgctgattc tggagttcga gccgacggag gcctgggaca
     1801 gcaccgggcc gcacgcgctg cgcaacgcga tgttcgcgct cgaaagtgcc cccaacctgc
     1861 gggcgctggc cgaggtggcg acccagacgg tccgcgagct gacgggcttt gaccgggtga
     1921 tgctctacaa atttgccccc gacgccaccg gcgaagtgat tgccgaggcc cgccgtgagg
     1981 ggctgcacgc ctttctgggc caccgttttc ccgcgtcgga cattccggcg caggcccgcg
     2041 cgctctacac ccggcacctg ctgcgcctga ccgccgacac ccgcgccgcc gccgtgccgc
     2101 tcgatcccgt cctcaacccg cagacgaatg cgcccacccc gctgggcggc gccgtgctgc
     2161 gcgccacctc gcccatgcac atgcagtacc tgcggaacat gggcgtcggg tcgagcctgt
     2221 cggtgtcggt ggtggtcggc ggccagctct ggggcctgat cgcctgccac caccagacgc
     2281 cctacgtgtt gccgcccgac ctgcgaacca cgctcgaata cctgggccgc ttgctgagcc
     2341 tgcaagttca ggtcaaggaa gcggcggacg tggcggcctt tcgccagagc ctgcgggagc
     2401 accacgcgcg ggtggccctc gcggcggcgc actcgctctc gccgcacgac accctcagtg
     2461 acccggcgct tgacctgctg ggcctgatgc gggccggggg cctgattctg cgtttcgagg
     2521 gccgctggca gacgttgggt gaagtgccgc ctgccccggc ggtggacgcg ctgctggcgt
     2581 ggctcgaaac ccagccgggc gccctggtcc agaccgacgc gctaggccaa ctgtggcccg
     2641 ccggcgccga tctcgccccc agcgcagcgg gcctgctcgc catcagcgtg ggcgagggct
     2701 ggtcggagtg cctcgtctgg ctgcggcccg aactgcggct ggaggtcgcc tggggcgggg
     2761 ccactcctga ccaggcgaaa gacgacctcg ggccgcgcca ctcattcgac acctacctcg
     2821 aagaaaaacg cggctacgcc gagccctggc atcccggcga aatcgaggag gcgcaggatc
     2881 tacgtgacac attgaccggg gcgctgggcg agtactttta tttctccatc gtctctgcgg
     2941 tggttggcat tctgctggtc gtggtcttgg gggtggtctt tgggatcctc tctagaaaca
     3001 aatgtggaca gaggagcaaa tttgggatca accgccctgc tgtgctggcg ccagaggatg
     3061 ggctggccat gtccctacac ttcatgacac tgggtggcag ttctctttcc cctactgagg
     3121 gcaaaggctc cggactccag ggccacatca tggagaaccc acagtacttc agtgatacct
     3181 gtgtccacca tatcaagcgc caggacatca ttctcaagtg ggagctaggg gagggagcct
     3241 ttggaaaggt ctttcttgct gagtgctaca accttctgaa tgatcaggac aagatgctgg
     3301 tggctgtcaa ggcactgaag gagacatctg agaatgctcg tcaggacttc catcgtgagg
     3361 cagagctgct caccatgcta cagcaccaac acatcgtacg cttctttgga gtctgcacgg
     3421 agggtggccc attgctcatg gtcttcgagt acatgcgcca tggggacctc aaccgtttcc
     3481 tccggtccca cggacctgat gcaaaactgc tggctggcgg cgaggatgtg gctcctggtc
     3541 ctttgggcct tgggcagctt ctggctgtgg ctagccaggt ggctgctggt atggtgtacc
     3601 tagccagcct gcactttgtg caccgggacc tggccacacg caactgtctg gtgggtcagg
     3661 gactagtggt caagattgga gacttcggca tgagcaggga catctacagc acagactact
     3721 accgtgtggg aggtcggacc atgctgccca tccgctggat gcctccagag agcatcctct
     3781 accgcaagtt cagcaccgag agtgatgtgt ggagcttcgg ggtggtgctc tgggagatct
     3841 tcacctatgg aaagcaaccc tggtaccagc tctccaacac tgaggcgatc gagtgcatca
     3901 cgcagggccg ggagctggag cggccgcgcg cctgccctcc tgatgtctac gccatcatgc
     3961 gcggctgctg gcagcgtgag ccgcaacagc gcctcagcat gaaggatgtg cacgcgcggc
     4021 tgcaggcctt ggcacaggcg ccaccgagtt acctggacgt tctgggctct agaggcggtg
     4081 gtggcagcgg aggagggggg agcggcggcg gtggcagtgg cggcggcggt agtcgcagcc
     4141 gctttgtgaa aaaagatagc gccggcagtg cgggtagtgc ggggagcgcc ggtagctatc
     4201 tggcgaacga aattctgtgg ggcagcgctg gtagcgcagg aagcgccggc agtgcagggt
     4261 tttgctatga aaacgaagtg gcgctgtcat aatcagccat accacatttg tagaggtttt
     4321 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4381 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4441 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4501 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4561 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4621 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4681 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4741 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4801 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4861 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4921 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4981 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5041 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5101 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5281 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5341 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5401 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5461 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5521 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5581 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5641 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5701 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5761 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5821 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5881 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5941 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6001 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     6061 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6121 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6181 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6241 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6301 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6361 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6421 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6481 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6541 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6601 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6661 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6721 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6841 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6901 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6961 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7021 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7081 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7141 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7201 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7261 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7321 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7381 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7441 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7501 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7561 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
