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
//
