LOCUS       BB2_L_BC_syn_BbsI_7      2215 bp    DNA     circular UNA 
DEFINITION  Note: the plasmid backbone was PCR amplified, only the linker sequence was checked by sequencing! (compare Seq folder)
FEATURES             Location/Qualifiers
     Vector          join(513..2215,1..408)
                     /origin="PCRproductNewBB2 (2.125bp circular)Target
                     vector: from AgeI cut at 5^6 to SpeI cut at 2116^2117,
                     2.111bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Region          124..181
                     /label="pUC origin"
     primer_bind     282..299
                     /label=pSTBlue_Seq_FW
     primer_bind     351..368
                     /label=BB2_seq_fw
     primer_bind     complement(391..407)
                     /label=BB2_BW
     Site            join(408..>408,complement(<409..>412),408^409,complement(408^409))
                     /site_type="restriction site"
                     /note="Name: SpeI"
                     /note="Pattern: actagt"
                     /note="cut_0_on_positive_strand: 2116^2117"
                     /note="cut_0_on_negative_strand: 2120^2121"
                     /note="inhibited_by: "
                     /note="star_activity: yes"
                     /note="site_type: other"
                     /note="restriction site"
                     /label=SpeI
     primer          409..453
                     /label=1_B1007_TT_BsaI
     primer          complement(413..460)
                     /label=2_B1007_TT_BsaI
     primer          454..483
                     /label=3_BsaI_FsB_Bam
     Region          454..459
                     /label=BsaI
     primer          complement(461..490)
                     /label=4_BsaI_FsB_BamXho
     FS              461..464
                     /label="FS B"
     FS              465..468
                     /label="FS 1"
     Region          complement(471..476)
                     /label=BbSI
     primer          484..512
                     /label=5_BsaI_FsC_Xho
     Region          489..494
                     /label=BbSI
     primer          complement(491..516)
                     /label=6_BsaI_FsC
     FS              497..500
                     /label="FS 4"
     FS              501..504
                     /label="FS C"
     Region          complement(506..511)
                     /label=BsaI
     Site            join(<513..>516,<517..517,512^513)
                     /site_type="restriction site"
                     /note="Name: AgeI"
                     /note="Pattern: accggt"
                     /note="cut_0_on_positive_strand: 5^6"
                     /note="cut_0_on_negative_strand: 9^10"
                     /note="inhibited_by: 5': 5-methylcytosine"
                     /note="site_type: other"
                     /note="restriction site"
                     /label=AgeI
     primer_bind     518..533
                     /label=BB2_FW
     Region          complement(<518..534)
                     /label="f1 origin"
     primer_bind     complement(609..628)
                     /label=BB2_seq_bw
     CDS             666..1523
                     /label=AmpicillinR
     misc_feature    1374..1390
                     /feature_type="Primer Region"
                     /label=BB2_BsaIMut_FW
     misc_feature    complement(1374..1390)
                     /feature_type="Primer Region"
                     /label=BB2_BsaIMut_BW
     primer          1382
                     /label="G changed to C"
ORIGIN
        1 GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT
       61 TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT
      121 GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC
      181 ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT
      241 GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG
      301 CGGAAGAGCG CCCAATACGC AAACCGCCTC TCCCCGCGCG TTGGCCGATT CATTAATGCA
      361 GCTGGCACGA CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGCAACACT AGTcgcaaaa
      421 aaccccgccc ctgacagggc ggggtttttt cgcGGTCTCC CCGGGGAGGC GTCTTCGGAT
      481 CCCTCGAGGA AGACGCCGCT AATTGGAGAC CACCGGTGCC GCTACAGGGC GCGTCAGGTG
      541 GCACTTTTCG GGGAAATGTG CGCGGAACCC CTATTTGTTT ATTTTTCTAA ATACATTCAA
      601 ATATGTATCC GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT TGAAAAAGGA
      661 AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG GCATTTTGCC
      721 TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA AGATGCTGAA GATCAGTTGG
      781 GTGCACGAGT GGGTTACATC GAACTGGATC TCAACAGCGG TAAGATCCTT GAGAGTTTTC
      841 GCCCCGAAGA ACGTTTTCCA ATGATGAGCA CTTTTAAAGT TCTGCTATGT GGCGCGGTAT
      901 TATCCCGTAT TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT TCTCAGAATG
      961 ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG ACAGTAAGAG
     1021 AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC GGCCAACTTA CTTCTGACAA
     1081 CGATCGGAGG ACCGAAGGAG CTAACCGCTT TTTTGCACAA CATGGGGGAT CATGTAACTC
     1141 GCCTTGATCG TTGGGAACCG GAGCTGAATG AAGCCATACC AAACGACGAG CGTGACACCA
     1201 CGATGCCTGT AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA CTACTTACTC
     1261 TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA GGACCACTTC
     1321 TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA ATCTGGAGCC GGTGAGCGTG
     1381 GCTCTCGCGG TATCATTGCA GCACTGGGGC CAGATGGTAA GCCCTCCCGT ATCGTAGTTA
     1441 TCTACACGAC GGGGAGTCAG GCAACTATGG ATGAACGAAA TAGACAGATC GCTGAGATAG
     1501 GTGCCTCACT GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT ATACTTTAGA
     1561 TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT TTTGATAATC
     1621 TCATGAACAA TAAAACTGTC TGCTTACATA AACAGTAATA CAAGGGGTGT AGATCTCATG
     1681 ACCAAAATCC CTTAACGTGA GTTTTCGTTC CACTGAGCGT CAGACCCCGT AGAAAAGATC
     1741 AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT GCTGCTTGCA AACAAAAAAA
     1801 CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC TACCAACTCT TTTTCCGAAG
     1861 GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTCC TTCTAGTGTA GCCGTAGTTA
     1921 GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC TCGCTCTGCT AATCCTGTTA
     1981 CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG GGTTGGACTC AAGACGATAG
     2041 TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT CGTGCACACA GCCCAGCTTG
     2101 GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG AGCTATGAGA AAGCGCCACG
     2161 CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG GCAGGGTCGG AACAG
//
