LOCUS Exported 6970 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses OptoOFF-TtCBD-mFGFR1 (light-inactivated receptor tyrosine kinase). ACCESSION . VERSION . KEYWORDS OptoOFF-TtCBD-mFGFR1_624 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6970 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..48) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 394..413 /label=T7 /note="T7 promoter, forward primer" promoter 394..412 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 466..475 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 472..513 /codon_start=1 /product="N-myristoylation signal from Src kinase (Pellman et al., 1985; Kaplan et al., 1988)" /label=myr /translation="MGSSKSKPKDPSQR" CDS 2443..2469 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2546..2563) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2552..2776 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2822..3250 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2909..2928) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3119..3140 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3259..3279) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3264..3593 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3444..3579 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3506..3525 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3660..4454 /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(3714..3733) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4324..4343 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4628..4749 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4665..4684) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4719..4738 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4798..4814) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4798..4814) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4811..4833) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4822..4838 /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(4846..4876) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4891..4912 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5029..5046) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5200..5788) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5280..5299) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5959..6819) /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 6582..6601 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6820..6924) /gene="bla" /label=AmpR promoter ORIGIN 1 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 61 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag 121 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt 181 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt 241 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 301 cataacttac ggtaaatggc ccgcctggga ggtctatata agcagagctc tctggctaac 361 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca 421 agctggctag cgtttaaacg ggccctctag actcgagcgg ccgcggccac catggggagt 481 agcaagagca agcctaagga ccccagccag cgcctcgaca tgaagagcgg caccaagaag 541 agcgacttcc atagccagat ggctgtgcac aagctggcca agagcatccc tctgcgcaga 601 caggtaacag tgtcagctga ctccagtgca tccatgaact ctggggttct cctggttcgg 661 ccctcacggc tctcctccag cgggaccccc atgctggctg gagtctccga atatgagctc 721 cctgaggatc cccgctggga gctgccacga gacagactgg tcttaggcaa accacttggc 781 gagggctgct tcgggcaggt ggtgttggct gaggccatcg ggctggataa ggacaaaccc 841 aaccgtgtga ccaaagtggc cgtgaagatg ttgaagtccg acgcaacgga gaaggacctg 901 tcggatctga tctcggagat ggagatgatg aaaatgattg ggaagcacaa gaatatcatc 961 aaccttctgg gagcgtgcac acaggatggt cctctttatg tcattgtgga gtacgcctcc 1021 aaaggcaatc tccgggagta tctacaggcc cggaggcctc ctgggctgga gtactgctat 1081 aaccccagcc acaaccccga ggaacagctg tcttccaaag atctggtatc ctgtgcctat 1141 caggtggctc ggggcatgga gtatcttgcc tctaagaagt gtatacaccg agacctggct 1201 gctaggaacg tcctggtgac cgaggataac gtaatgaaga tcgcagactt tggcttagct 1261 cgagacattc atcatatcga ctactacaag aaaaccacca acggccggct gcctgtgaag 1321 tggatggccc ctgaggcgtt gtttgaccgg atctacacac accagagcga tgtgtggtct 1381 tttggagtgc tcttgtggga gatcttcact ctgggtggct ccccataccc cggtgtgcct 1441 gtggaggaac ttttcaagct gctgaaggag ggtcatcgaa tggacaagcc cagtaactgt 1501 accaatgagc tgtacatgat gatgcgggac tgctggcatg cagtgccctc tcagagacct 1561 acgttcaagc agttggtgga agacctggac cgcattgtgg ccttgacctc caaccaggag 1621 tatctggacc tgtccatacc gctggaccag tactcaccca gctttcccga cacacggagc 1681 tccacctgct cctcagggga ggactctgtc ttctctcatg agccgttacc tgaggagccc 1741 tgtctgcctc gacaccccac ccagcttgcc aacagtggac tcaaacggcg cgtcgagacc 1801 ggtcccgagg acctcggcac cggactcctc gaagcactct tgagaggaga tttggccggc 1861 gccgaggctc tctttcgacg ggggcttagg ttttggggac ccgagggtat tctggagcac 1921 ctgctcctgc ctgtgcttcg ggaggtggga gaagcctggc atcgcggcga gatcggtgtg 1981 gccgaggagc atctggcatc cacatttctg cgcgcgagac tgcaggagct gctcgacctc 2041 gccgggttcc cacctggccc ccccgtgctg gtaaccacgc caccagggga gaggcacgag 2101 atcggcgcaa tgttggctgc ctatcacctg cgaaggaagg gcgtgccagc gctgtacttg 2161 ggaccagaca cccccctccc cgatctcaga gcacttgcga gacggctcgg agccggagcg 2221 gtggttctgt cagctttgct ttccgagcct ctcagggcgt tgccagacgg cgcactgaaa 2281 gacttggcac ctcgggtgtt cctgggaggg caaggagccg gccctgaaga ggcccgacgg 2341 ctcggggccg agtacatgga agatctgaag ggattggccg aagcactgtg gcttccaaga 2401 ggaccagaga aagaagcaat ccccggtgga tccggagtcg actatccgta cgacgtacca 2461 gactacgcac tcgactaaga attccaccac actggactag tggatccgag ctcggtacca 2521 agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 2581 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2641 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 2701 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 2761 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc 2821 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 2881 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 2941 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 3001 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 3061 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 3121 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 3181 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 3241 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 3301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3421 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3481 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 3541 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 3601 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 3661 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3721 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3781 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3841 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3901 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3961 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 4021 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 4081 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 4141 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 4201 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4261 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4321 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4381 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4441 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4621 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 4681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 4741 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 4801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 4861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 4921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 4981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 5401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5641 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 6001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 6061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 6121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 6181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 6241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 6961 atcgggagat //