LOCUS Exported 6967 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses OptoOFF-MxCBD-mFGFR1 (light-inactivated receptor tyrosine kinase). ACCESSION . VERSION . KEYWORDS OptoOFF-MxCBD-mFGFR1_623 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6967) 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 6967) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6967 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 409..428 /label=T7 /note="T7 promoter, forward primer" promoter 409..427 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 481..490 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 487..528 /codon_start=1 /product="N-myristoylation signal from Src kinase (Pellman et al., 1985; Kaplan et al., 1988)" /label=myr /translation="MGSSKSKPKDPSQR" CDS 2458..2484 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2561..2578) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2567..2791 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2837..3265 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2924..2943) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3134..3155 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3274..3294) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3279..3608 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3459..3594 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3521..3540 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3675..4469 /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(3729..3748) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4339..4358 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4643..4764 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4680..4699) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4734..4753 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4813..4829) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4813..4829) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4826..4848) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4837..4853 /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(4861..4891) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4906..4927 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5044..5061) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5215..5800) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5295..5314) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5971..6831) /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 6594..6613 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6832..6936) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tgggaggtct atataagcag 361 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca 421 ctatagggag acacaagctg gctagcgttt aaacgggccc tctagactcg agcggccgcg 481 gccaccatgg ggagtagcaa gagcaagcct aaggacccca gccagcgcct cgacatgaag 541 agcggcacca agaagagcga cttccatagc cagatggctg tgcacaagct ggccaagagc 601 atccctctgc gcagacaggt aacagtgtca gctgactcca gtgcatccat gaactctggg 661 gttctcctgg ttcggccctc acggctctcc tccagcggga cccccatgct ggctggagtc 721 tccgaatatg agctccctga ggatccccgc tgggagctgc cacgagacag actggtctta 781 ggcaaaccac ttggcgaggg ctgcttcggg caggtggtgt tggctgaggc catcgggctg 841 gataaggaca aacccaaccg tgtgaccaaa gtggccgtga agatgttgaa gtccgacgca 901 acggagaagg acctgtcgga tctgatctcg gagatggaga tgatgaaaat gattgggaag 961 cacaagaata tcatcaacct tctgggagcg tgcacacagg atggtcctct ttatgtcatt 1021 gtggagtacg cctccaaagg caatctccgg gagtatctac aggcccggag gcctcctggg 1081 ctggagtact gctataaccc cagccacaac cccgaggaac agctgtcttc caaagatctg 1141 gtatcctgtg cctatcaggt ggctcggggc atggagtatc ttgcctctaa gaagtgtata 1201 caccgagacc tggctgctag gaacgtcctg gtgaccgagg ataacgtaat gaagatcgca 1261 gactttggct tagctcgaga cattcatcat atcgactact acaagaaaac caccaacggc 1321 cggctgcctg tgaagtggat ggcccctgag gcgttgtttg accggatcta cacacaccag 1381 agcgatgtgt ggtcttttgg agtgctcttg tgggagatct tcactctggg tggctcccca 1441 taccccggtg tgcctgtgga ggaacttttc aagctgctga aggagggtca tcgaatggac 1501 aagcccagta actgtaccaa tgagctgtac atgatgatgc gggactgctg gcatgcagtg 1561 ccctctcaga gacctacgtt caagcagttg gtggaagacc tggaccgcat tgtggccttg 1621 acctccaacc aggagtatct ggacctgtcc ataccgctgg accagtactc acccagcttt 1681 cccgacacac ggagctccac ctgctcctca ggggaggact ctgtcttctc tcatgagccg 1741 ttacctgagg agccctgtct gcctcgacac cccacccagc ttgccaacag tggactcaaa 1801 cggcgcgtcg agaccggtcc acacgcagag acttggaggg aatctatgct tgccgccaca 1861 caagcctacg accagcctag agtatcagat gtactggatg aggtccttgc ggctctgccc 1921 cctctgaagg ccttcgatga agtgctggcc cctttgctgt gcgatgtcgg agagcggtgg 1981 gagagcggaa ccctgacagt tgcgcaggaa catctggtct cacagatggt gcgcgcccgg 2041 ctggtgagtc tgctgcacgc ggcaccattg ggacgccaca gacatggcgt tctcgcctgt 2101 ttcccagagg aggagcatga gatgggcttg cttggtgccg ccttgagact ccgccatctc 2161 ggcgttagag taaccctgct cggccagcga gtgccagccg aggacctcgg gcgagcagtg 2221 ttggccctgc gcccggactt cgtgggcctg tcaacagttg caagcaggag cgcagaggac 2281 ttcgaggata ccttgacccg actccgccag gccctgccaa ggggcctccc tgtatgggtg 2341 ggcggggcag ccgcaaggtc tcatcaggcc gtgtgcgagc gcctggcagt ccatgttttt 2401 cagggcgaag aagattggga tagacttgcc ggaacacccg gtggatccgg agtcgactat 2461 ccgtacgacg taccagacta cgcactcgac taagaattcc accacactgg actagtggat 2521 ccgagctcgg taccaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt 2581 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 2641 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 2701 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc 2761 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 2821 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2881 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2941 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 3001 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3061 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3121 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3181 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 3241 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3301 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3361 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3421 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 3481 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3541 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 3601 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 3661 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3721 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3781 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3841 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3901 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3961 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4021 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4081 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4141 atgatctgga cgaagaacat caggggctcg cgccagccga actgttcgcc aggctcaagg 4201 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4261 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4321 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagaactt ggcggcgaat 4381 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4441 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 4501 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4561 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4621 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 4681 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 4741 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 4801 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4861 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4921 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 4981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttgcgc 5041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 5461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 5521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 5581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5641 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 5881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 5941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gggacccacg 6121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 6481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 6541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 6601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 6661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 6721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 6781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 6841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 6901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 6961 tgacgtc //