LOCUS BB3_hygro_ARSCEN_URA3_PT_MAL509 4040 bp DNA circular UNA COMMENT Freezer software: ID 37277 FEATURES Location/Qualifiers Vector 1..678 /origin="pSTBLUE-1 wo_ORI_circular_pUC_ORI_aus_pGAPZB (2.745bp circular)Target vector: from BbsI cut at 2054^2055 to BbsI cut at 2732^2733, 678bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" Vector join(1941..4040,1..722) /origin="MST607_BB3_hygro_ARSCEN_pUC-ORI_linkerAD (2.852bp circular)Target vector: from BsaI cut at 752^753 to BsaI cut at 722^723, 2.822bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" oriT complement(5..678) /label="pUC_ORI_aus_pGAPZB" misc_feature join(<1941..1944,679..>722) /feature_type="Insertion" /origin="FS1FsA_BX_FsDFS2_BW (106bp linear)Fragment: from BbsI cut at 12^13 to BbsI cut at 90^91, 78bpOrientation: reverse complementedPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" terminator 683..722 /label="BBa_B1007" misc_feature join(<1857..>1860,<723..>726) /feature_type="Insertion" /origin="BsaI_A_1_4_B_FW (118bp linear)Fragment: from SpeI cut at 5^6 to AgeI cut at 109^110, 104bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" misc_feature 723..1860 /feature_type="Insertion" /origin="BB2_A_1_4_B_FW_URA3_PCR_prod_Bpi1_FS1_4 (3.313bp circular)Fragment: from BsaI cut at 2163^2164 to BsaI cut at 3301^3302, 1.138bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" Vector join(1857..>1860,<723..726) /origin="PCRproductNewBB2_BsaI_A_1_4_B_FW (2,215bp circular)Target vector: from BpiI cut at 2199^2200 to BpiI cut at 2167^2168, 2,183bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" primer <727..745 /label="URA3_marker_ampl_fw1_BpiI_FS1" misc_feature 727..1856 /feature_type="Insertion" /origin="URA3_CBS7962_PT_PCR_prod_Bpi1_FS1_4 (1,162bp linear)Fragment: from BpiI cut at 14^15 to BpiI cut at 1144^1145, 1,130bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" promoter 731..963 /label="URA3 promoter (pRS vectors)" gene 964..1767 /score="0.99" /source="AUGUSTUS" /ID="g2024" /Product="Ura3p" /Note="corresponds to s288c YEL021W" primer 1215..1249 /label="URA3_marker_mut_fw2" primer complement(1215..1249) /label="URA3_marker_mut_rev2" Site 1233 /site_type="mutagenized site" /label="A->G" primer 1621..1650 /label="URA3_marker_mut_fw3" primer complement(1621..1650) /label="URA3_marker_mut_rev3" Site 1635 /site_type="mutagenized site" /label="C->G" terminator 1768..1856 /label="Terminator Region" primer complement(1844..>1860) /label="URA3_marker_ampl_rev1_BpiI_FS4" misc_feature <1861..>1900 /feature_type="Insertion" /origin="BsaI_B_1_4_C_FW (118bp linear)Fragment: from SpeI cut at 5^6 to AgeI cut at 109^110, 104bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" misc_feature 1861..1900 /feature_type="Insertion" /origin="BB2_BsaI_B_1_4_C_FW (2.215bp circular)Fragment: from BsaI cut at 2163^2164 to BsaI cut at 2203^2204, 40bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" Region complement(1871..1876) /label="BbSI" Region 1889..1894 /label="BbSI" misc_feature <1901..>1940 /feature_type="Insertion" /origin="BsaI_C_1_4_D_FW (118bp linear)Fragment: from SpeI cut at 5^6 to AgeI cut at 109^110, 104bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" misc_feature 1901..1940 /feature_type="Insertion" /origin="BB2_BsaI_C_1_4_D_FW (2.215bp circular)Fragment: from BsaI cut at 2163^2164 to BsaI cut at 2203^2204, 40bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" misc_feature <1945..>3524 /feature_type="Insertion" /origin="hphMX4_wo_BspMI (1.600bp linear)Fragment: full sequenceOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" misc_feature 1945..3524 /feature_type="Insertion" /origin="pSTBLUE-1 wo_Kan_circular_hphMX4_wo_BspMI (4.635bp circular)Fragment: from BbsI cut at 863^864 to BbsI cut at 2443^2444, 1.580bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" CDS 1949..2292 /label="TEFPromoter" misc_feature 2122 /label="C to G" CDS 2293..3321 /label="hphR" misc_feature 2604 /label="G to C" CDS 3330..3524 /label="TEFterminator" rep_origin <3525..4040 /label="CEN ARS6" misc_feature 3525..4040 /feature_type="Insertion" /origin="BB1_FS3_CEN ARS 6_FS4 (3.452bp circular)Fragment: from BbsI cut at 2149^2150 to BbsI cut at 2665^2666, 516bpOrientation: originalPlus strand 5': no changePlus strand 3': no changeMinus strand 5': no changeMinus strand 3': no change" /label="Insert" ORIGIN 1 CGCTGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT 61 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT 121 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC 181 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA 241 GCGTGGCGCT TTCTCAATGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT 301 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA 361 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG 421 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC 481 CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA 541 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG 601 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT 661 TGATCTTTTC TACGGGGTGG AGcgcaaaaa accccgcccc tgacagggcg gggttttttc 721 gcGATCGGAG CAGGGTCCAT AAAGCTTTTC AATTCATCTT TTTTTTTTTA TTCTTTTTTT 781 TGATTCCGGT TTCCTTGAAA TTTTTTTGAT TCGGTAATCT CCGAGCAGAA GGAAGAACGA 841 AGGAAGGAGC ACAGACTTAG ATTGGTATAT ATACGCATAT GTAGTGTTGA AGAAACATGA 901 AATTGCCCAG TATTCTTAAC CCAACTGCAC AGAACAAAAA CCTGCAGGAA ACGAAGATAA 961 ATCATGTCGA AAGCTACATA TAAGGAACGT GCTGCTACTC ATCCTAGTCC TGTTGCTGCC 1021 AAGCTATTTA ATATCATGCA CGAAAAGCAA ACAAACTTGT GTGCTTCATT GGATGTTCGT 1081 ACCACCAAGG AATTACTGGA GTTAGTTGAA GCATTAGGTC CCAAAATTTG TTTACTAAAA 1141 ACACATGTGG ATATCTTGAC TGATTTTTCC ATGGAGGGCA CAGTTAAGCC GCTAAAGGCA 1201 TTATCCGCCA AGTACAATTT TTTACTCTTC GAgGACAGAA AATTTGCTGA CATTGGTAAT 1261 ACAGTCAAAT TGCAGTACTC TGCGGGTGTA TACAGAATAG CAGAATGGGC AGACATTACG 1321 AATGCACACG GTGTGGTGGG CCCAGGTATT GTTAGCGGTT TGAAGCAGGC GGCGGAAGAA 1381 GTAACAAAGG AACCTAGAGG CCTTTTGATG TTAGCAGAAT TGTCATGCAA GGGCTCCCTA 1441 GCTACTGGAG AATATACTAA GGGTACTGTT GACATTGCGA AGAGCGACAA AGATTTTGTT 1501 ATCGGCTTTA TTGCTCAAAG AGACATGGGT GGAAGAGATG AAGGTTACGA TTGGTTGATT 1561 ATGACACCCG GTGTGGGTTT AGATGACAAG GGAGACGCAT TGGGTCAACA GTATAGAACC 1621 GTGGATGATG TGGTgTCTAC AGGATCTGAC ATTATTATTG TTGGAAGAGG ACTATTTGCA 1681 AAGGGAAGGG ATGCTAAGGT AGAGGGTGAA CGTTACAGAA AAGCAGGCTG GGAAGCATAT 1741 TTGAGAAGAT GCGGCCAGCA AAACTAAAAA ACTGTATTAT AAGTAAATGC ATGTATACTA 1801 AACTCACAAA TTAGAGCTTC AATTTAATTA TATCAGTTAT TACCCGGGAA TCTCGGCGCT 1861 CCGGGGAGGC GTCTTCGGAT CCCTCGAGGA AGACGCCGCT AATTGGAGGC GTCTTCGGAT 1921 CCCTCGAGGA AGACGCCGCT AGCTCATGGA CATGGAGGCC CAGAATACCC TCCTTGACAG 1981 TCTTGACGTG CGCAGCTCAG GGGCATGATG TGACTGTCGC CCGTACATTT AGCCCATACA 2041 TCCCCATGTA TAATCATTTG CATCCATACA TTTTGATGGC CGCACGGCGC GAAGCAAAAA 2101 TTACGGCTCC TCGCTGCAGA CGTGCGAGCA GGGAAACGCT CCCCTCACAG ACGCGTTGAA 2161 TTGTCCCCAC GCCGCGCCCC TGTAGAGAAA TATAAAAGGT TAGGATTTGC CACTGAGGTT 2221 CTTCTTTCAT ATACTTCCTT TTAAAATCTT GCTAGGATAC AGTTCTCACA TCACATCCGA 2281 ACATAAACAA CCATGGGTAA AAAGCCTGAA CTCACCGCGA CGTCTGTCGA GAAGTTTCTG 2341 ATCGAAAAGT TCGACAGCGT CTCCGACCTG ATGCAGCTCT CGGAGGGCGA AGAATCTCGT 2401 GCTTTCAGCT TCGATGTAGG AGGGCGTGGA TATGTCCTGC GGGTAAATAG CTGCGCCGAT 2461 GGTTTCTACA AAGATCGTTA TGTTTATCGG CACTTTGCAT CGGCCGCGCT CCCGATTCCG 2521 GAAGTGCTTG ACATTGGGGA ATTCAGCGAG AGCCTGACCT ATTGCATCTC CCGCCGTGCA 2581 CAGGGTGTCA CGTTGCAAGA CCTCCCTGAA ACCGAACTGC CCGCTGTTCT GCAGCCGGTC 2641 GCGGAGGCCA TGGATGCGAT CGCTGCGGCC GATCTTAGCC AGACGAGCGG GTTCGGCCCA 2701 TTCGGACCGC AAGGAATCGG TCAATACACT ACATGGCGTG ATTTCATATG CGCGATTGCT 2761 GATCCCCATG TGTATCACTG GCAAACTGTG ATGGACGACA CCGTCAGTGC GTCCGTCGCG 2821 CAGGCTCTCG ATGAGCTGAT GCTTTGGGCC GAGGACTGCC CCGAAGTCCG GCACCTCGTG 2881 CACGCGGATT TCGGCTCCAA CAATGTCCTG ACGGACAATG GCCGCATAAC AGCGGTCATT 2941 GACTGGAGCG AGGCGATGTT CGGGGATTCC CAATACGAGG TCGCCAACAT CTTCTTCTGG 3001 AGGCCGTGGT TGGCTTGTAT GGAGCAGCAG ACGCGCTACT TCGAGCGGAG GCATCCGGAG 3061 CTTGCAGGAT CGCCGCGGCT CCGGGCGTAT ATGCTCCGCA TTGGTCTTGA CCAACTCTAT 3121 CAGAGCTTGG TTGACGGCAA TTTCGATGAT GCAGCTTGGG CGCAGGGTCG ATGCGACGCA 3181 ATCGTCCGAT CCGGAGCCGG GACTGTCGGG CGTACACAAA TCGCCCGCAG AAGCGCGGCC 3241 GTCTGGACCG ATGGCTGTGT AGAAGTACTC GCCGATAGTG GAAACCGACG CCCCAGCACT 3301 CGTCCGAGGG CAAAGGAATA ATCAGTACTG ACAATAAAAA GATTCTTGTT TTCAAGAACT 3361 TGTCATTTGT ATAGTTTTTT TATATTGTAG TTGTTCTATT TTAATCAAAT GTTAGCGTGA 3421 TTTATATTTT TTTTCGCCTC GACATCATCT GCCCAGATGC GAAGTTAAGT GCGCAGAAAG 3481 TAATATCATG CGTCAATCGT ATGTGAATGC TGGTCGCTAT ACTGGCTTCC TTTTCATCAC 3541 GTGCTATAAA AATAATTATA ATTTAAATTT TTTAATATAA ATATATAAAT TAAAAATAGA 3601 AAGTAAAAAA AGAAATTAAA GAAAAAATAG TTTTTGTTTT CCGAAGATGT AAAAGACTCT 3661 AGGGGGATCG CCAACAAATA CTACCTTTTA TCTTGCTCTT CCTGCTCTCA GGTATTAATG 3721 CCGAATTGTT TCATCTTGTC TGTGTAGAAc ACCACACACG AAAATCCTGT GATTTTACAT 3781 TTTACTTATC GTTAATCGAA TGTATATCTA TTTAATCTGC TTTTCTTGTC TAATAAATAT 3841 ATATGTAAAG TACGCTTTTT GTTGAAATTT TTTAAACCTT TGTTTATTTT TTTTTCTTCA 3901 TTCCGTAACT CTTCTACCTT CTTTATTTAC TTTCTAAAAT CCAAATACAA AACATAAAAA 3961 TAAATAAACA CAGAGTAAAT TCCCAAATTA TTCCATCATT AAAAGATACG AGGCGCGTGT 4021 AAGTTACAGG CAAGCGATCG //