LOCUS       BB2_L_CD_syn_BbsI_8      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
     Region          join(2213..2215,1..55)
                     /label="pUC origin"
     Vector          join(387..2215,1..282)
                     /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"
     primer_bind     156..173
                     /label=pSTBlue_Seq_FW
     primer_bind     225..242
                     /label=BB2_seq_fw
     primer_bind     complement(265..281)
                     /label=BB2_BW
     Site            join(282..>282,complement(<283..>286),282^283,complement(282^283))
                     /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          283..327
                     /label=1_B1007_TT_BsaI
     primer          complement(287..334)
                     /label=2_B1007_TT_BsaI
     primer          328..357
                     /label=3_BsaI_FsC_Bam
     Region          328..333
                     /label=BsaI
     primer          complement(335..364)
                     /label=4_BsaI_FsC_BamXho
     FS              335..338
                     /label="FS C"
     FS              339..342
                     /label="FS 1"
     primer          358..386
                     /label=5_BsaI_FsD_Xho
     primer          complement(365..390)
                     /label=6_BsaI_FsD
     FS              371..374
                     /label="FS 4"
     FS              375..378
                     /label="FS D"
     Region          complement(380..385)
                     /label=BsaI
     Site            join(<387..>390,<391..391,386^387)
                     /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     392..407
                     /label=BB2_FW
     Region          complement(<392..408)
                     /label="f1 origin"
     primer_bind     complement(483..502)
                     /label=BB2_seq_bw
     CDS             540..1397
                     /label=AmpicillinR
     misc_feature    1248..1264
                     /feature_type="Primer Region"
                     /label=BB2_BsaIMut_FW
     misc_feature    complement(1248..1264)
                     /feature_type="Primer Region"
                     /label=BB2_BsaIMut_BW
     primer          1256
                     /label="G changed to C"
ORIGIN
        1 ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT GCTCACATGT
       61 TCTTTCCTGC GTTATCCCCT GATTCTGTGG ATAACCGTAT TACCGCCTTT GAGTGAGCTG
      121 ATACCGCTCG CCGCAGCCGA ACGACCGAGC GCAGCGAGTC AGTGAGCGAG GAAGCGGAAG
      181 AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC
      241 ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA CACTAGTcgc aaaaaacccc
      301 gcccctgaca gggcggggtt ttttcgcGGT CTCCAATTGG AGGCGTCTTC GGATCCCTCG
      361 AGGAAGACGC CGCTAGCTGG AGACCACCGG TGCCGCTACA GGGCGCGTCA GGTGGCACTT
      421 TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT CTAAATACAT TCAAATATGT
      481 ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA ATATTGAAAA AGGAAGAGTA
      541 TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG
      601 TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG TTGGGTGCAC
      661 GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT CCTTGAGAGT TTTCGCCCCG
      721 AAGAACGTTT TCCAATGATG AGCACTTTTA AAGTTCTGCT ATGTGGCGCG GTATTATCCC
      781 GTATTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG
      841 TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT
      901 GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG ACAACGATCG
      961 GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG GGATCATGTA ACTCGCCTTG
     1021 ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA CGAGCGTGAC ACCACGATGC
     1081 CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT
     1141 CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT
     1201 CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG CGTGGCTCTC
     1261 GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC CCGTATCGTA GTTATCTACA
     1321 CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA GATCGCTGAG ATAGGTGCCT
     1381 CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT
     1441 TAAAACTTCA TTTTTAATTT AAAAGGATCT AGGTGAAGAT CCTTTTTGAT AATCTCATGA
     1501 ACAATAAAAC TGTCTGCTTA CATAAACAGT AATACAAGGG GTGTAGATCT CATGACCAAA
     1561 ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA
     1621 TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG
     1681 CTACCAGCGG TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT
     1741 GGCTTCAGCA GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC
     1801 CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG
     1861 GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG
     1921 GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA
     1981 ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC
     2041 GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG
     2101 AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC
     2161 TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAA
//
