LOCUS Exported 4379 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial ST1 repression YFP reporter: protospacer B. ACCESSION . VERSION . KEYWORDS SK-YFP-ST1-B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4379) AUTHORS Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM TITLE Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. JOURNAL Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. PUBMED 24076762 REFERENCE 2 (bases 1 to 4379) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4379 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(163..191) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 786..806 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 908..994 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(1529..2479) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 2527..2749 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator 3241..3335 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3366..4160) /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(3477..3496) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4087..4106 /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4339..4357 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 gatccgataa atatctaaca ccgtgcgtgt tgactatttt acctctggcg gtgataatgg 61 ttgcatgtac tagaattctt tctttaagaa ggagatatac atatgaattc ttcatggcga 121 atacttttaa agtctcgtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt 181 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat 241 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca 301 tggccaacac ttgtcactac tttcggttat ggtctaaaat gctttgctag atacccagat 361 catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga 421 actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt 481 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt 541 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa 601 caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt 661 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca 721 gacaaccatt acctgtccta tcaatctgcc ctttcgaaag atcccaacga aaagagagac 781 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta 841 tacaaataag cttaaccgaa gcgtttgata gttgatatcc tttgcctgcg gccgcaacta 901 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg 961 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg 1021 ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc 1081 cggcttcagg tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc 1141 cacgggaggc gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc 1201 ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat 1261 ttcatgttct agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg 1321 ttcatctgtt acattgtcga tctgttcatg gtgaacagct ttaaatgcac caaaaactcg 1381 taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt 1441 tccctttgat atctaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc 1501 actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct 1561 ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatgcttac 1621 tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag 1681 catcgtgtag tgtttttctt agtccgttac gtaggtagga atctgatgta atggttgttg 1741 gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt 1801 gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca 1861 ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt 1921 taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa 1981 ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact 2041 cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta 2101 tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact aattctaatt 2161 tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag 2221 gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat 2281 aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg 2341 tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa 2401 ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga 2461 aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat 2521 tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta 2581 aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg 2641 ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga 2701 aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt 2761 cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac 2821 cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc 2881 cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct 2941 gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt 3001 catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt 3061 atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct 3121 gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc 3181 acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg 3241 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag 3301 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctctcga accccagagt 3361 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg 3421 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata 3481 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg 3541 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg 3601 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct 3661 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc 3721 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga 3781 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca 3841 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc 3901 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat 3961 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa 4021 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc 4081 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc 4141 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat 4201 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc 4261 ccaaccttac cagagggcgc cccagctggc aattccgacg tctaagaaac cattattatc 4321 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca cctcgaggg //