LOCUS Exported 6985 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression vector for DTXK-lumitoxin. ACCESSION . VERSION . KEYWORDS pcDNA3.1-DTXK-lumitoxin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6985) AUTHORS Schmidt D, Tillberg PW, Chen F, Boyden ES TITLE A fully genetically encoded protein architecture for optical control of peptide ligand concentration. JOURNAL Nat Commun. 2014 Jan 10;5:3019. doi: 10.1038/ncomms4019. PUBMED 24407101 REFERENCE 2 (bases 1 to 6985) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6985 /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" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 946..969 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1165..1188 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1678..1824 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" CDS 1834..2544 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1979..1997) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2247..2266 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2576..2593) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2582..2806 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2852..3280 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2939..2958) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3149..3170 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3289..3309) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3294..3623 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3474..3609 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3536..3555 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3690..4484 /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(3744..3763) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4354..4373 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4658..4779 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4695..4714) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4749..4768 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4828..4844) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4828..4844) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4841..4863) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4852..4868 /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(4876..4906) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4921..4942 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5059..5076) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5230..5818) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5310..5329) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5989..6849) /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 6612..6631 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6850..6954) /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 tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggccgacta caaggacgac 961 gacgacaagc tgaagcttgc cgccaagtac tgcaagctgc ccctgagaat cggtccatgc 1021 aagcggaaga tccccagctt ctactacaag tggaaggcca agcagtgcct gcccttcgac 1081 tactctggct gcggaggcaa cgccaaccgg ttcaagacca tcgaggaatg cagacggacc 1141 tgcgtgggag gtaccgccgc cgccgactac aaggacgacg acgacaagat cgacgccgcc 1201 gccggcggcg ccctgtgcaa cgaattcttg gctactacac ttgaacgtat tgagaagaac 1261 tttgtcatta ctgacccaag attgccagat aatcccatta tattcgcgtc cgatagtttc 1321 ttgcagttga cagaatatag ccgtgaagaa attttgggaa gaaactgcag gtttctacaa 1381 ggtcctgaaa ctgatcgcgc gacagtgaga aaaattagag atgccataga taaccaaaca 1441 gaggtcactg ttcagctgat taattataca aagagtggta aaaagttctg gaacctcttt 1501 cacttgcagc ctatgcgaga tcagaaggga gatgtccagt actttattgg ggttcagttg 1561 gatggaactg agcatgtccg agatgctgcc gagagagagg gagtcatgct gattaagaaa 1621 actgcagaaa atattgatga ggcggcaaaa gaacttagcg gccgcggtac tcgagttgct 1681 gtgggccagg acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg 1741 gtgatctcag ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc 1801 atgctttggc agaagaaacc acgtaggatt cgtatggtga gcaagggcga ggaggataac 1861 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc 1921 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 1981 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag 2041 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag 2101 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg 2161 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg 2221 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag 2281 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg 2341 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag 2401 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac 2461 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc 2521 ggcatggacg agctgtacaa gtaatctaga gggcccgttt aaacccgctg atcagcctcg 2581 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 2641 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 2701 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 2761 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 2821 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 3001 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 3061 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 3181 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 3301 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3361 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3421 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3541 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 3601 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 3661 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3721 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3781 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3841 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3901 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3961 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 4021 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 4081 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4141 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4201 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4261 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4321 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4381 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4441 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4501 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4561 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4621 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 4681 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 4741 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 4801 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 4861 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4921 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4981 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 5041 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 5101 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5161 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5221 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5281 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 5341 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5401 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 5461 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 5521 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 5581 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 5641 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 5701 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 5761 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5821 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5881 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5941 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6001 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6061 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6121 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6961 tccccgaaaa gtgccacctg acgtc //