LOCUS Exported 2164 bp ds-DNA circular SYN 05-DEC-2018 DEFINITION MoClo golden gate assembly AB part for pL-lacO1 (LacI-repressed promoter, as used in DOI:10.1038/35002125).. ACCESSION . VERSION . KEYWORDS P30m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2164) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 2164) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 5, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers promoter 5..60 /ApEinfo_fwdcolor="#80ff80" /ApEinfo_revcolor="#80ffff" /Color="#ffffff" /modified_by="Andrey" /Transferred_From="P30m - PL-lacO1" /Transferred_Similarity="100.00%" /label="PL-lacO1" protein_bind 9..25 /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).; color: #31849b" /X-Transferred_From="lac_operator" /X-Transferred_Similarity="88.24%" /Transferred_From="P30m - PL-lacO1" /Transferred_Similarity="100.00%" /label="lac operator" promoter 25..60 /ApEinfo_fwdcolor="#80ff80" /ApEinfo_revcolor="#80ffff" /X-Transferred_From="P30m - PL-lacO1" /X-Transferred_Similarity="100.00%" /modified_by="Andrey" /Transferred_From="pLlacO-1" /Transferred_Similarity="100.00%" /label="PL-lacO1 sigma70 promoter" protein_bind 31..47 /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).; color: #31849b" /X-Transferred_From="lac_operator" /X-Transferred_Similarity="100.00%" /Transferred_From="P30m - PL-lacO1, lac_operator" /Transferred_Similarity="100.00%" /label="lac operator" misc_feature 65..2164 /Color="#cc99ff" /Transferred_From="CIDAR_DVA_backbone" /Transferred_Similarity="100.00%" /label="CIDAR DVA backbone" misc_feature complement(66..71) /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 72..92 /Color="#a6ccff" /Transferred_From="BioBrick_suffix" /Transferred_Similarity="100.00%" /label="BioBrick suffix" primer_bind 2060..2080 /Sequence="AAATAGGCGTATCACGAGGCA" /Hairpin_Tm="46.1" /Tm="59.6" /Self_Dimer_Tm="2.4" /%GC="47.6" /created_by="primer3" /Transferred_From="Part.Seq.FOR" /Transferred_Similarity="100.00%" /label="Part.Seq.FOR" terminator complement(2083..2126) /Color="#ffffff" /Transferred_From="bacterial_terminator" /Transferred_Similarity="100.00%" /label="bacterial terminator" misc_feature 2129..2150 /Color="#99ccff" /Transferred_From="BioBrick_prefix_(2)" /Transferred_Similarity="100.00%" /label="BioBrick prefix" misc_feature 2129..2148 /Color="#99ccff" /Transferred_From="BioBrick_prefix" /Transferred_Similarity="100.00%" /label="BioBrick prefix" misc_feature 2158..2163 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 2158..2163 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" source 1..2164 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 10..24 /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)." protein_bind 31..47 /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)." misc_feature 72..92 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(98..115) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(269..857) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(349..368) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1028..1888) /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 1651..1670 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1889..1993) /gene="bla" /label=AmpR promoter primer_bind 2061..2079 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2083..2126 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 2129..2150 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 ggagataaat gtgagcggat aacattgaca ttgtgagcgg ataacaagat actgagcaca 61 tactagagac ctactagtag cggccgctgc aggcttcctc gctcactgac tcgctgcgct 121 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 181 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 241 accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct gacgagcatc 301 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 361 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 421 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 481 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 541 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 601 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 661 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 721 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 781 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 841 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 901 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 961 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 1021 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 1081 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 1141 ctggccccag tgctgcaatg ataccgcgcg acccacgctc accggctcca gatttatcag 1201 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 1261 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 1321 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 1381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 1441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 1501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 1561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 1621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 1681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 1741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 1801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 1861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 1921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 1981 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta 2041 tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat 2101 ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag actagtgggt 2161 ctca //