LOCUS Exported 8932 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of mCherry-Syk in mammalian cells. ACCESSION . VERSION . KEYWORDS pMSCV-mCherry-Syk SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8932) AUTHORS Strijbis K, Tafesse FG, Fairn GD, Witte MD, Dougan SK, Watson N, Spooner E, Esteban A, Vyas VK, Fink GR, Grinstein S, Ploegh HL TITLE Bruton's Tyrosine Kinase (BTK) and Vav1 contribute to Dectin1-dependent phagocytosis of Candida albicans in macrophages. JOURNAL PLoS Pathog. 2013 Jun;9(6):e1003446. doi: 10.1371/journal.ppat.1003446. Epub 2013 Jun 27. PUBMED 23825946 REFERENCE 2 (bases 1 to 8932) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8932 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 1416..1425 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1422..2129 /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(1567..1585) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1835..1854 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2029..2052 /label=DsRed1-C /note="DsRed1, forward primer" promoter 4075..4574 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(4110..4133) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4507..4526 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4595..5194 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4595..4614) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5091..5111 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5294..5808 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" primer_bind complement(5977..5993) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5977..5993) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5990..6012) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6001..6017 /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(6025..6055) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6070..6091 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6208..6225) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6379..6967) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6459..6478) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7138..7998) /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 7761..7780 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7999..8103) /gene="bla" /label=AmpR promoter primer_bind 8171..8189 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8227..8249) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8349..8368 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8562..8584 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8712..8731 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 8931..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatctgccac catggtgagc aagggcgagg 1441 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg 1501 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc 1561 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt 1621 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact 1681 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg 1741 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg 1801 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg 1861 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca 1921 agcagaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca 1981 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ctggacatca 2041 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact 2101 ccaccggcgg catggacgag ctgtacaagg gaggaggagg atcaggagga ggaggatcac 2161 ctagggcggg aagtgctgtg gacagcgcca accacctgac ctactttttt ggcaacatca 2221 cccgggaaga ggctgaagac tacctggtcc agggaggcat gaccgatggg ctctacctgc 2281 tacgccagag ccgcaattac ctgggtggtt ttgctttgtc ggtggctcac aacaggaagg 2341 cacaccacta caccatcgag agggaactta atggcaccta cgccatctcc gggggcaggg 2401 cccatgccag cccagcagac ctctgccatt accactccca ggaacctgat ggccttatct 2461 gcctccttaa gaagcccttc aaccggcccc cgggagtaca gcccaagacc ggaccctttg 2521 aggacctgaa ggagaacctc atcagggaat atgtgaaaca gacctggaac cttcagggcc 2581 aggctctgga gcaagccatc atcagccaga agccccagct ggagaagctg atcgccacca 2641 cggcccatga gaagatgccc tggttccatg gcaacatctc cagagatgaa tcagagcaga 2701 cggtcctcat agggtcaaag accaatggaa aattcctgat cagggccaga gacaacagcg 2761 gctcctatgc tctgtgcctg ctgcacgaag ggaaagtatt gcactaccgc attgacaggg 2821 acaagaccgg gaagctctcc attcctgagg ggaagaagtt tgacaccctc tggcagctag 2881 tggaacatta ctcttacaag ccagatgggc tactaagagt cctcacggta ccatgccaaa 2941 agattggtgc acagatgggc cacccaggaa gcccaaatgc ccatcccgtg acttggtcac 3001 cgggtggaat aatctcaagg atcaaatcct actccttccc aaagcctggc cacaaaaagc 3061 ctgccccacc ccaagggagc cgtccagaga gcactgtgtc cttcaacccc tatgagccaa 3121 cgggagggcc ctggggccca gacagaggcc ttcagagaga agccctgccc atggacacag 3181 aggtgtacga gagcccttat gctgaccctg aagagatccg gcccaaagag gtctacctgg 3241 acaggagcct gctgaccctg gaggacaatg aactgggctc cggtaacttc gggactgtga 3301 aaaagggata ctaccaaatg aaaaaagttg tgaaaaccgt ggctgtgaaa atcctgaaga 3361 acgaggccaa cgacccggct ttgaaggacg agctgctggc agaggcgaac gtcatgcagc 3421 agctggacaa cccctacatt gtgcgcatga tcggaatctg cgaggcggag tcctggatgc 3481 tggtgatgga gatggcggag ctggggccgc tcaacaagta cctgcagcag aacaggcaca 3541 ttaaggataa gaacatcata gagctggttc accaggtttc catggggatg aagtatttgg 3601 aagagagcaa ctttgtgcac agagatctgg ctgcgcggaa cgtgcttctg gtcacacagc 3661 actatgccaa gatcagcgat ttcggtcttt ccaaagccct gcgtgctgat gaaaactact 3721 acaaggccca gacccacggg aagtggcccg tgaagtggta cgcccccgaa tgcatcaact 3781 actacaagtt ctccagtaag agtgacgtct ggagcttcgg agtcctgatg tgggaagcgt 3841 tctcctatgg gcagaagccc tacagaggga tgaaagggag cgaagtgacc gccatgctgg 3901 agaaaggaga gcggatgggg tgccctgcag gatgcccgag agagatgtac gacctgatga 3961 acctgtgctg gacttacgat gtggagaaca ggccaggatt cacagctgtg gaactgaggc 4021 ttcgcaatta ctactacgac gtggttaact aactcgaggt taacgaattc taccgggtag 4081 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt 4141 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac 4201 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt 4261 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca 4321 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca 4381 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt 4441 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag 4501 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat 4561 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg 4621 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga 4681 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct 4741 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 4801 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 4861 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 4921 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 4981 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 5041 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 5101 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg 5161 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc 5221 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca 5281 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc 5341 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga 5401 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg 5461 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga 5521 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac 5581 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa 5641 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac 5701 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg 5761 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt 5821 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca 5881 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag 5941 cgcagaagag ctggggagaa ttaattcgta atcatggtca tagctgtttc ctgtgtgaaa 6001 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 6061 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 6121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 6181 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 6241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 6301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 6361 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 6421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 6481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6541 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 6601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6781 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 6841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 7021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 7081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 7141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 7201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 7261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 7321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 7381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 7441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 7501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 8041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 8101 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 8161 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg 8221 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc 8281 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct 8341 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc 8401 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa 8461 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg 8521 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 8581 aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg 8641 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga 8701 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg 8761 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata 8821 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac 8881 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aa //