LOCUS Exported 4347 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Co-expresses Syt1-SNARE37aa-linker complex with p045. ACCESSION . VERSION . KEYWORDS pDC184 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4347) AUTHORS Zhou Q, Lai Y, Bacaj T, Zhao M, Lyubimov AY, Uervirojnangkoorn M, Zeldin OB, Brewster AS, Sauter NK, Cohen AE, Soltis SM, Alonso-Mori R, Chollet M, Lemke HT, Pfuetzner RA, Choi UB, Weis WI, Diao J, Sudhof TC, Brunger AT TITLE Architecture of the synaptotagmin-SNARE machinery for neuronal exocytosis. JOURNAL Nature. 2015 Sep 3;525(7567):62-7. doi: 10.1038/nature14975. Epub 2015 Aug 17. PUBMED 26280336 REFERENCE 2 (bases 1 to 4347) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4347 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 3..27 /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)." CDS 86..115 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 116..136 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" promoter 403..421 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 422..446 /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)." CDS 705..749 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" primer_bind complement(787..805) /label=T7 Term /note="T7 terminator, reverse primer" terminator 801..848 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(1068..1727) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 1642..1661 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(1728..1830) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind complement(2222..2239) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2356..2900) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(2436..2455) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3125..4207) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 4169..4188 /label=LacI-R /note="LacI, reverse primer" promoter complement(4208..4285) /gene="lacI" /label=lacI promoter promoter 4331..2 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag 61 gagatatacc atggcatcgt actaccatca ccatcaccat caccatcacc atcacgaaaa 121 cctgtatttt cagggcagca acaggagact gcagcagacc caggcccagg tggatgaggt 181 ggtggacatc atgagggtga atgtggacaa ggtcctggag cgagaccaga agctatcgga 241 actggatgat cgcgcagatg ccctccaggc aggggcctcc cagtttgaaa caagtgcagc 301 caagctcaag cgcaaatact ggtagctcga caagcttgcg gccgcataat gcttaagtcg 361 aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag 421 gggaattgtg agcggataac aattccccat cttagtatat tagttaagta taagaaggag 481 atatacatat ggccctcagt gagatcgaga ccaggcacag tgagatcatc aagttggaga 541 acagcatccg ggagctacac gatatgttca tggacatggc catgctggtg gagagccagg 601 gggagatgat tgacaggatc gagtacaatg tggaacacgc tgtggactac gtggagaggg 661 ccgtgtctga caccaagaag gccgtcaagt aactcgagtc tggtaaagaa accgctgctg 721 cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg 781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg 841 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca 901 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga 961 cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca 1021 ggcgtagcac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc 1081 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca 1141 cagacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa 1201 tatttgccca tagtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca 1261 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct 1321 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga 1381 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca 1441 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt 1501 gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga 1561 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg 1621 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc 1681 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta 1741 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg 1801 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag 1861 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc 1921 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt 1981 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc 2041 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag 2101 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca 2161 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt 2221 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg 2281 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc 2341 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg 2401 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttcccctggc 2461 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta 2521 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa 2581 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta 2641 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa 2701 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa 2761 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga 2821 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc 2881 gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt 2941 cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt 3001 aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct 3061 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt 3121 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3181 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca 3241 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc 3301 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga 3361 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc 3421 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca 3481 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca 3541 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt 3601 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg 3661 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat 3721 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa 3781 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga 3841 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc 3901 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa 3961 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca 4021 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg 4081 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca 4141 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg 4201 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa 4261 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc 4321 attaggaaat taatacgact cactata //