LOCUS Exported 2192 bp ds-DNA circular SYN 05-DEC-2018 DEFINITION MoClo golden gate assembly BC part for BCD24 (low expression bi-cistronic RBS, engineered for downstream context-independence; see https://doi.org/10.1038/nmeth.2404).. ACCESSION . VERSION . KEYWORDS U8m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2192) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 2192) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 5, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers misc_feature 1..2192 /Original_Bases /label /note="Geneious type: Editing History Insertion" rbs 5..88 /created_by="murraylab" /label="BCD24" misc_feature 93..2192 /ApEinfo_revcolor="#6ecff6" /ApEinfo_fwdcolor="#6ecff6" /label="DVLA" misc_feature complement(94..99) /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" primer_bind complement(176..196) /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 2088..2108 /Sequence="AAATAGGCGTATCACGAGGCA" /Hairpin_Tm="46.5" /Tm="59.6" /Self_Dimer_Tm="None" /%GC="47.6" /created_by="primer3" /label="SC189/Part.Seq.FOR" misc_feature 2186..2191 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 2186..2191 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" source 1..2192 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 100..120 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(126..143) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(297..885) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(377..396) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1056..1916) /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 1679..1698 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1917..2021) /gene="bla" /label=AmpR promoter primer_bind 2089..2107 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2111..2154 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 2157..2178 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 tactgggccc aagttcactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa 61 catcttaatc atgcgatgga cggtttctaa tgagagacct actagtagcg gccgctgcag 121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 301 cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 721 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 961 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 1021 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 1081 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 1141 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgcgac 1201 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 1261 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 1321 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 1381 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 1441 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 1501 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 1561 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 1621 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 1681 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 1741 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 1801 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 1861 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 1921 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 1981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 2041 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc 2101 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat 2161 tcgcggccgc ttctagagac tagtgggtct ca //