LOCUS Exported 6686 bp ds-DNA circular SYN 12-AUG-2021 DEFINITION Active SNAP tag at the centrosome. ACCESSION . VERSION . KEYWORDS pCS2-mCherry_SNAP_SNAPV_PACT_Active SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6686) AUTHORS Bucko PJ, Lombard CK, Rathbun L, Garcia I, Bhat A, Wordeman L, Smith FD, Maly DJ, Hehnly H, Scott JD TITLE Subcellular drug targeting illuminates local kinase action. JOURNAL Elife. 2019 Dec 24;8. pii: 52220. doi: 10.7554/eLife.52220. PUBMED 31872801 REFERENCE 2 (bases 1 to 6686) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6686 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..978 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1013..1031 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1013..1030 /label=SP6 /note="SP6 promoter, forward primer" regulatory 1059..1068 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1065..1772 /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(1210..1228) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1478..1497 /label=mCherry-F /note="mCherry, forward primer" CDS 1791..2333 /codon_start=1 /product="human O6-alkylguanine-DNA-alkyltransferase variant that forms covalent bonds with benzylguanine derivatives" /label=SNAP-tag(R) /note="optimized for mammalian expression" /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEV PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV VKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLAV KEWLLAHEGHRLGKPGLG" promoter complement(3694..3711) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 3716..3850 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3740..3759) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3794..3813 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3963..3983) /label=T3 /note="T3 promoter, forward primer" promoter complement(3963..3981) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4002..4018) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4002..4018) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4015..4037) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4026..4042 /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(4050..4080) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4095..4116 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4233..4250) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4404..4992) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4484..4503) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5163..6023) /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 5786..5805 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6024..6128) /gene="bla" /label=AmpR promoter rep_origin complement(6154..6609) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6291..6312) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6503..6522 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt 61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca 121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt 181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg 241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat 301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca 361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt 421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac 481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac 541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta 601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa 661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg 721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac 781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg 841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa 901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc 961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt 1021 gacactatag aatacaagct acttgttctt tttgcagtgc caccatggtg agcaagggcg 1081 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct 1141 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca 1201 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc 1261 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg 1321 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg 1381 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca 1441 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca 1501 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga 1561 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct 1621 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca 1681 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc 1741 actccaccgg cggcatggac gagctgtaca agggtactgg ctctggaacc gacaaagact 1801 gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa gctggaactg tctgggtgcg 1861 aacagggcct gcacgagatc aagctgctgg gcaaaggaac atctgccgcc gacgccgtgg 1921 aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc actgatgcag gccaccgcct 1981 ggctcaacgc ctactttcac cagcctgagg ccatcgagga gttccctgtg ccagccctgc 2041 accacccagt gttccagcag gagagcttta cccgccaggt gctgtggaaa ctgctgaaag 2101 tggtgaagtt cggagaggtc atcagctacc agcagctggc cgccctggcc ggcaatcccg 2161 ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc cgtgcccatt ctgatcccct 2221 gccaccgggt ggtgtctagc tctggcgccg tggggggcta cgagggcggg ctcgccgtga 2281 aagagtggct gctggcccac gagggccaca gactgggcaa gcctgggctg ggtgggagtg 2341 gaacaggcag cggtaccggc tcagataagg actgcgagat gaaacggacc acactggact 2401 caccactggg gaaactggag ctgagcggct gcgagcaggg actccacgaa atcaaactgc 2461 tcggcaaggg gactagcgcc gctgatgctg tggaggtccc agctcctgca gccgtcctgg 2521 gaggacccga acccctcatg caagctaccg catggctgaa cgcatacttc catcagccag 2581 aagctatcga agaatttccc gtgcccgccc tccaccatcc cgtctttcag caagaaagtt 2641 tcacacggca ggtcctctgg aagctcctga aggtggtcaa attcggcgaa gtgatctcat 2701 atcagcaact cgctgcactg gcaggaaacc ctgcagctac cgcagcagtg aagacagctc 2761 tgtctgggaa ccccgtccct atcctgattc cttgccatag agtggtcagc tccagcggag 2821 cagtgggagg atatgaggga ggactggctg tcaaggaatg gctcctggca catgaaggcc 2881 ataggctggg aaaacccgga ctgggacccg ccgggattgg agcacctggg agcctcgaga 2941 cgggttctgg gacaggatcg ggctctgctt ggaagccaga cgaaacggct ccacagagtt 3001 ccctgaggcg cccagacccc ggccggcttc caccagctgc cagcgaggaa gcacacacca 3061 gcaatgtcaa gatggaaaaa ttgtacctgc attacttgag agcagagagc tttagaaaag 3121 ctctgattta tcaaaagaag tatcttttgc tgttgattgg tggattccag gattctgaac 3181 aagaaacact ctccatgatt gcccatttgg gggtatttcc ttccaaagca gaacggaaaa 3241 tcacatctcg tcctttcacc aggttccgca cggccgtcag ggtggtcatt gcaatattaa 3301 gattacgttt tttggttaag aaatggcaag aagtagatcg gaaaggagct ctggcacaag 3361 gcaaagcccc tcgcccaggg ccccgagcac gacagccgca gtctccaccc agaaccagag 3421 agtccccccc aacccgggat gtaccctctg gccacaccag ggaccctgcc agaggccgca 3481 gactggcagc agcagcctcc ccacacagtg ggggaagagc cactccatcc ccaaattcaa 3541 gattagaaag atccctgact gcttctcaag atccagaaca ttccttgaca gagtatattc 3601 accatttaga agtgatccag caaagattgg gaggggtact accagattct acttcaaaga 3661 aatcctgcca cccgatgaac ctgcagtcta gaactatagt gagtcgtatt acgtagatcc 3721 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa 3781 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 3841 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg 3901 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt 3961 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg 4021 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa 4081 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 4141 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 4201 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 4261 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 4321 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 4381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 4441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4561 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 4621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 4741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 4801 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 4861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 4921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 4981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5041 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 5101 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 5161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 5221 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 5281 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 5341 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 5401 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 5461 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 5521 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 5581 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 5641 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 5701 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 5761 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 5821 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 5881 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 5941 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6001 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6061 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 6121 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa 6181 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 6241 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 6301 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 6361 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc 6421 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 6481 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 6541 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 6601 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg 6661 cctcttcgct attacgccag tcgacc //