LOCUS       sequence-367691-        7837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optically controlled eDrcMet expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7837)
  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 7837)
  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: eDrMet
FEATURES             Location/Qualifiers
     source          1..7837
                     /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    4625..4746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4662..4681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4716..4735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4753..5208)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4890..4911)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5102..5121
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5235..5339
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5341..5698
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5341..5532
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5364..5384)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5549..5684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5733..6527
                     /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             5733..6527
                     /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(5787..5806)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6397..6416
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6715..6734)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6759..6806
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7135..7723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7624..7643
                     /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 tctagaaaaa
     3001 agagaaagca aattaaagat ctgggcagtg aattagttcg ctacgatgca agagtacaca
     3061 ctcctcattt ggataggctt gtaagtgccc gaagtgtaag cccaactaca gaaatggttt
     3121 caaatgaatc tgtagactac cgagctactt ttccagaaga tcagtttcct aattcatctc
     3181 agaacggttc atgccgacaa gtgcagtatc ctctgacaga catgtccccc atcctaacta
     3241 gtggggactc tgatatatcc agtccattac tgcaaaatac tgtccacatt gacctcagtg
     3301 ctctaaatcc agagctggtc caggcagtgc agcatgtagt gattgggccc agtagcctga
     3361 ttgtgcattt caatgaagtc ataggaagag ggcattttgg ttgtgtatat catgggactt
     3421 tgttggacaa tgatggcaag aaaattcact gtgctgtgaa atccttgaac agaatcactg
     3481 acataggaga agtttcccaa tttctgaccg agggaatcat catgaaagat tttagtcatc
     3541 ccaatgtcct ctcgctcctg ggaatctgcc tgcgaagtga agggtctccg ctggtggtcc
     3601 taccatacat gaaacatgga gatcttcgaa atttcattcg aaatgagact cataatccaa
     3661 ctgtaaaaga tcttattggc tttggtcttc aagtagccaa aggcatgaaa tatcttgcaa
     3721 gcaaaaagtt tgtccacaga gacttggctg caagaaactg tatgctggat gaaaaattca
     3781 cagtcaaggt tgctgatttt ggtcttgcca gagacatgta tgataaagaa tactatagtg
     3841 tacacaacaa aacaggtgca aagctgccag tgaagtggat ggctttggaa agtctgcaaa
     3901 ctcaaaagtt taccaccaag tcagatgtgt ggtcctttgg cgtgctcctc tgggagctga
     3961 tgacaagagg agccccacct tatcctgacg taaacacctt tgatataact gtttacttgt
     4021 tgcaagggag aagactccta caacccgaat actgcccaga ccccttatat gaagtaatgc
     4081 taaaatgctg gcaccctaaa gccgaaatgc gcccatcctt ttctgaactg gtgtcccgga
     4141 tatcagcgat cttctctact ttcattgggg agcactatgt ccatgtgaac gctacttatg
     4201 tgaacgtaaa atgtgtcgct ccgtatcctt ctctgttgtc atcagaagat aacgctgatg
     4261 atgaggtgga cacacgacca gcctccttct gggagacatc atctagaggc ggtggtggca
     4321 gcggaggagg ggggagcggc ggcggtggca gtggcggcgg cggtagtcgc agccgctttg
     4381 tgaaaaaaga tagcgccggc agtgcgggta gtgcggggag cgccggtagc tatctggcga
     4441 acgaaattct gtggggcagc gctggtagcg caggaagcgc cggcagtgca gggttttgct
     4501 atgaaaacga agtggcgctg tcataatcag ccataccaca tttgtagagg ttttacttgc
     4561 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4621 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4741 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4801 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4861 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4921 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4981 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5041 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5101 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5161 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5221 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5281 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5341 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5401 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5461 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5521 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5581 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5641 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5701 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5761 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5821 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5881 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5941 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6001 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6061 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6121 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6181 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6241 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6301 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6361 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6421 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6481 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6541 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6601 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6661 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6721 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6781 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6841 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6901 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6961 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     7021 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7081 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7141 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7201 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7261 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7321 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7381 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7441 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7501 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7561 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7621 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7681 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7741 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7801 ccctgattct gtggataacc gtattaccgc catgcat
//
