LOCUS Exported 2210 bp ds-DNA circular SYN 05-DEC-2018 DEFINITION MoClo golden gate assembly DE part for gN2 gRNA (guide RNA for S. pyogenes Cas9; sequence designed with NUPACK for stable tracrRNA structure and open targeting region).. ACCESSION . VERSION . KEYWORDS UC20m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2210) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 2210) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 5, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers misc_RNA 5..24 /created_by="sclamons" /label="gN2 target" misc_feature <25..66 /Source="referenceToDocument: urn:local:.:h9-9u0o4st" /label="Digestion of IA_UC15 extraction (concatenated sequence 2)" /note="Geneious type: Concatenated sequence" misc_RNA 25..66 /created_by="User" /label="SP tracrRNA constant region" terminator 67..106 /created_by="User" /label="tracrRNA Terminator" misc_feature <67..>106 /Source="referenceToDocument: urn:local:.:ha-9u0o4st" /label="Digestion of IA_T22 extraction (concatenated sequence 3)" /note="Geneious type: Concatenated sequence" misc_feature 107^108 /Original_Bases="AT" /label="AT" /note="Geneious type: Editing History Deletion" misc_feature 109..110 /Original_Bases /label /note="Geneious type: Editing History Insertion" misc_feature 111..2210 /ApEinfo_revcolor="#6ecff6" /ApEinfo_fwdcolor="#6ecff6" /label="DVLA" primer_bind complement(194..214) /Sequence="TTACCGCCTTTGAGTGAGCTG" /Hairpin_Tm="None" /Tm="60.6" /Self_Dimer_Tm="None" /%GC="52.4" /created_by="primer3" /label="SC190/Part.Seq.REV" primer_bind 2106..2126 /Sequence="AAATAGGCGTATCACGAGGCA" /Hairpin_Tm="46.5" /Tm="59.6" /Self_Dimer_Tm="None" /%GC="47.6" /modified_by="User" /label="SC189/Part.Seq.FOR" source 1..2210 /organism="synthetic DNA construct" /mol_type="other DNA" misc_RNA 25..100 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 118..138 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(144..161) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(315..903) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(395..414) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1074..1934) /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 1697..1716 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1935..2039) /gene="bla" /label=AmpR promoter primer_bind 2107..2125 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2129..2172 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 2175..2196 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 tactctcctc ctccccccca atatgtttta gagctagaaa tagcaagtta aaataaggct 61 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttaggt agagacctac 121 tagtagcggc cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 301 gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc 361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 1021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 1081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 1141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 1201 gcaatgatac cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 1261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 1321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 1381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 1441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 1501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 1561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 1621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 1681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 1741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 1801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 1861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 1921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 1981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 2041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc 2101 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct 2161 aaggatgatt tctggaattc gcggccgctt ctagagacta gtgggtctca //