LOCUS       sequence-209199-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OptoOFF-MxCBD-mFGFR1 (light-inactivated receptor tyrosine 
            kinase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  AUTHORS   Kainrath S, Stadler M, Reichhart E, Distel M, Janovjak H
  TITLE     Green-Light-Induced Inactivation of Receptor Signaling Using 
            Cobalamin-Binding Domains.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Apr 10;56(16):4608-4611. doi: 
            10.1002/anie.201611998. Epub 2017 Mar 20.
  PUBMED    28319307
REFERENCE   2  (bases 1 to 6970)
  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: 
            "10.1002/anie.201611998"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2017-04-10- 10"; volume: "56"; issue: "16"; pages: 
            "4608-4611"
COMMENT     Sequence Label: OptoOFF-MxCBD-mFGFR1_623
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     188..207
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        188..206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      260..269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             266..307
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             2237..2263
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2340..2357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2346..2570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2616..3044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2703..2722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2913..2934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3053..3073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3058..3387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3238..3373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3300..3319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3454..4248
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3508..3527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4118..4137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4422..4543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4459..4478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4513..4532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4592..4608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4592..4608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4605..4627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4616..4632
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4640..4646,4647..4664,4665..4670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4685..4706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4823..4840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4994..5582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5074..5093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5753..6544,6545..6613))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6376..6395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6614..6718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6793..6812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      181 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta aacgggccct
      241 ctagactcga gcggccgcgg ccaccatggg gagtagcaag agcaagccta aggaccccag
      301 ccagcgcctc gacatgaaga gcggcaccaa gaagagcgac ttccatagcc agatggctgt
      361 gcacaagctg gccaagagca tccctctgcg cagacaggta acagtgtcag ctgactccag
      421 tgcatccatg aactctgggg ttctcctggt tcggccctca cggctctcct ccagcgggac
      481 ccccatgctg gctggagtct ccgaatatga gctccctgag gatccccgct gggagctgcc
      541 acgagacaga ctggtcttag gcaaaccact tggcgagggc tgcttcgggc aggtggtgtt
      601 ggctgaggcc atcgggctgg ataaggacaa acccaaccgt gtgaccaaag tggccgtgaa
      661 gatgttgaag tccgacgcaa cggagaagga cctgtcggat ctgatctcgg agatggagat
      721 gatgaaaatg attgggaagc acaagaatat catcaacctt ctgggagcgt gcacacagga
      781 tggtcctctt tatgtcattg tggagtacgc ctccaaaggc aatctccggg agtatctaca
      841 ggcccggagg cctcctgggc tggagtactg ctataacccc agccacaacc ccgaggaaca
      901 gctgtcttcc aaagatctgg tatcctgtgc ctatcaggtg gctcggggca tggagtatct
      961 tgcctctaag aagtgtatac accgagacct ggctgctagg aacgtcctgg tgaccgagga
     1021 taacgtaatg aagatcgcag actttggctt agctcgagac attcatcata tcgactacta
     1081 caagaaaacc accaacggcc ggctgcctgt gaagtggatg gcccctgagg cgttgtttga
     1141 ccggatctac acacaccaga gcgatgtgtg gtcttttgga gtgctcttgt gggagatctt
     1201 cactctgggt ggctccccat accccggtgt gcctgtggag gaacttttca agctgctgaa
     1261 ggagggtcat cgaatggaca agcccagtaa ctgtaccaat gagctgtaca tgatgatgcg
     1321 ggactgctgg catgcagtgc cctctcagag acctacgttc aagcagttgg tggaagacct
     1381 ggaccgcatt gtggccttga cctccaacca ggagtatctg gacctgtcca taccgctgga
     1441 ccagtactca cccagctttc ccgacacacg gagctccacc tgctcctcag gggaggactc
     1501 tgtcttctct catgagccgt tacctgagga gccctgtctg cctcgacacc ccacccagct
     1561 tgccaacagt ggactcaaac ggcgcgtcga gaccggtcca cacgcagaga cttggaggga
     1621 atctatgctt gccgccacac aagcctacga ccagcctaga gtatcagatg tactggatga
     1681 ggtccttgcg gctctgcccc ctctgaaggc cttcgatgaa gtgctggccc ctttgctgtg
     1741 cgatgtcgga gagcggtggg agagcggaac cctgacagtt gcgcaggaac atctggtctc
     1801 acagatggtg cgcgcccggc tggtgagtct gctgcacgcg gcaccattgg gacgccacag
     1861 acatggcgtt ctcgcctgtt tcccagagga ggagcatgag atgggcttgc ttggtgccgc
     1921 cttgagactc cgccatctcg gcgttagagt aaccctgctc ggccagcgag tgccagccga
     1981 ggacctcggg cgagcagtgt tggccctgcg cccggacttc gtgggcctgt caacagttgc
     2041 aagcaggagc gcagaggact tcgaggatac cttgacccga ctccgccagg ccctgccaag
     2101 gggcctccct gtatgggtgg gcggggcagc cgcaaggtct catcaggccg tgtgcgagcg
     2161 cctggcagtc catgtttttc agggcgaaga agattgggat agacttgccg gaacacccgg
     2221 tggatccgga gtcgactatc cgtacgacgt accagactac gcactcgact aagaattcca
     2281 ccacactgga ctagtggatc cgagctcggt accaagctta agtttaaacc gctgatcagc
     2341 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2401 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2461 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2521 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2581 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2941 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3361 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3421 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3481 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3541 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3601 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3661 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3721 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3781 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3841 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3901 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3961 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4021 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4081 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4141 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4201 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4261 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4321 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4381 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4441 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4501 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4561 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4681 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4801 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4861 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4921 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5521 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5581 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5641 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5701 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5821 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5881 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5941 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6001 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6061 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6361 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6721 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6781 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6841 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6901 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     6961 tgtacgggcc
//
