LOCUS       sequence-367693-        8266 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Optically controlled eDrHER2 expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8266)
  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 8266)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: eDrHER2
FEATURES             Location/Qualifiers
     source          1..8266
                     /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..659
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     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    5054..5175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5091..5110)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5145..5164
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5182..5637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5319..5340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5531..5550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5664..5768
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5770..6127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5770..5961
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5793..5813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5978..6113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6162..6956
                     /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             6162..6956
                     /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(6216..6235)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6826..6845
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7144..7163)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7188..7235
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7564..8152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8053..8072
                     /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 agcagctcga gtccatcgtc tctgcggtgg
     2941 ttggcattct gctggtcgtg gtcttggggg tggtctttgg gatcctcatc aagcgacggc
     3001 agcagaagat ccggaagtac acgatgcgga gactgctgca ggaaacggag ctggtggagc
     3061 cgctgacacc tagcggagcg atgcccaacc aggcgcagat gcggatcctg aaagagacgg
     3121 agctgaggaa ggtgaaggtg cttggatctg gcgcttttgg cacagtctac aagggcatct
     3181 ggatccctga tggggagaat gtgaaaattc cagtggccat caaagtgttg agggaaaaca
     3241 catcccccaa agccaacaaa gaaatcttag acgaagcata cgtgatggct ggtgtgggct
     3301 ccccatatgt ctcccgcctt ctgggcatct gcctgacatc cacggtgcag ctggtgacac
     3361 agcttatgcc ctatggctgc ctcttagacc atgtccggga aaaccgcgga cgcctgggct
     3421 cccaggacct gctgaactgg tgtatgcaga ttgccaaggg gatgagctac ctggaggatg
     3481 tgcggctcgt acacagggac ttggccgctc ggaacgtgct ggtcaagagt cccaaccatg
     3541 tcaaaattac agacttcggg ctggctcggc tgctggacat tgacgagaca gagtaccatg
     3601 cagatggggg caaggtgccc atcaagtgga tggcgctgga gtccattctc cgccggcggt
     3661 tcacccacca gagtgatgtg tggagttatg gtgtgactgt gtgggagctg atgacttttg
     3721 gggccaaacc ttacgatggg atcccagccc gggagatccc tgacctgctg gaaaaggggg
     3781 agcggctgcc ccagcccccc atctgcacca ttgatgtcta catgatcatg gtcaaatgtt
     3841 ggatgattga ctctgaatgt cggccaagat tccgggagtt ggtgtctgaa ttctcccgca
     3901 tggccaggga cccccagcgc tttgtggtca tccagaatga ggacttgggc ccagccagtc
     3961 ccttggacag caccttctac cgctcactgc tggaggacga tgacatgggg gacctggtgg
     4021 atgctgagga gtatctggta ccccagcagg gcttcttctg tccagaccct gccccgggcg
     4081 ctgggggcat ggtccaccac aggcaccgca gctcatctac caggagtggc ggtggggacc
     4141 tgacactagg gctggagccc tctgaagagg aggcccccag gtctccactg gcaccctccg
     4201 aaggggctgg ctccgatgta tttgatggtg acctgggaat gggggcagcc aaggggctgc
     4261 aaagcctccc cacacatgac cccagccctc tacagcggta cagtgaggac cccacagtac
     4321 ccctgccctc tgagactgat ggctacgttg cccccctgac ctgcagcccc cagcctgaat
     4381 atgtgaacca gccagatgtt cggccccagc ccccttcgcc ccgagagggc cctctgcctg
     4441 ctgcccgacc tgctggtgcc actctggaaa ggcccaagac tctctcccca gggaagaatg
     4501 gggtcgtcaa agacgttttt gcctttgggg gtgccgtgga gaaccccgag tacttgacac
     4561 cccagggagg agctgcccct cagccccacc ctcctcctgc cttcagccca gccttcgaca
     4621 acctctatta ctgggaccag gacccaccag agcggggggc tccacccagc accttcaaag
     4681 ggacacctac ggcagagaac ccagagtacc tgggtctgga cgtgccagtg tctagaggcg
     4741 gtggtggcag cggaggaggg gggagcggcg gcggtggcag tggcggcggc ggtagtcgca
     4801 gccgctttgt gaaaaaagat agcgccggca gtgcgggtag tgcggggagc gccggtagct
     4861 atctggcgaa cgaaattctg tggggcagcg ctggtagcgc aggaagcgcc ggcagtgcag
     4921 ggttttgcta tgaaaacgaa gtggcgctgt cataatcagc cataccacat ttgtagaggt
     4981 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5041 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5101 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5161 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5221 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5281 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5341 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5401 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5461 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5521 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5581 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     5641 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5701 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5761 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     5821 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5881 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5941 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6001 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6061 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6121 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     6181 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6241 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6301 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     6361 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6421 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6481 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6541 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6601 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6661 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     6721 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6781 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6841 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6901 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6961 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     7021 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7081 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     7141 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     7201 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     7261 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     7321 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     7381 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     7441 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7501 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7561 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7621 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7681 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7741 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7801 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7861 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7921 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7981 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8041 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8101 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8161 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8221 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
