5' annealing region: atgagcggtggcgatggac

3' annealing region: gatattcatcggggtaagtaa

3' reverse complement: ttacttaccccgatgaatatc
5' oligo-F: ccaaaAGATCTatgagcggtggcgatggac
3' oligo-R: gctagCTCGAGttaGGATCCttacttaccccgatgaatatc
PCR ceaB-F/ceaB-R on CE2CEALIB (1777 bp, pcrpdt)
Digest pcrpdt (BglII/XhoI, 1761+11+5 b
p, L, pcrdig)
Digest pBca9145-Bca1089 (BglII/XhoI, 2057+696 bp, L, vec
tdig)
Ligate pcrdig and vectdig (pBca9145-ceaB)
-----------------------------------------
>ceaB-F Cloning of ceaB
ccaaaAGATCTatgagcggtggcgatggac
>ceaB-R Cloning of ceaB
gctagCTCGAGttaGGATCCttacttaccccgatgaatatc
>CE2CEALIB
aactcggttttaatcagacctggcatgagtggaagcgggacgaacagcacaggcaacaacaacgccgccccgggcacttc
cggggcatgagtatgtgatatccggggctgcaccccggaccccgccaacacatcacgggccacaaaattttttgtggccc
gctctgcgttttctaagtgttatccctcctgatttctaaaaaattttccacctgaacttgacagaaaaaacgatgacggg
tactttttgatctgtacataaaaccagtggttttatgtacagtattaatcatgtaattaattgttttaacgcttaaaaga
gggaatttttatgagcggtggcgatggacgcggccataacacgggcgcgcatagcacaagtggtaacattaatggtggcc
cgaccgggcttggtgtaggtggtggtgcttctgatggctccggatggagttcggaaaataacccgtggggtggtggttcc
ggtagcggcattcactggggtggtggttccggtcatggtaatggcggggggaatggtaattccggtggtggttcgggaac
aggcggtaatctgtcagcagtagctgcgccagtggcatttggttttccggcactttccactccaggagctggcggtctgg
cggtcagtatttcagcgggagcattatcggcagctattgctgatattatggctgccctgaaaggaccgtttaaatttggt
ctttggggggtggctttatatggtgtattgccatcacaaatagcgaaagatgaccccaatatgatgtcaaagattgtgac
gtcattacccgcagatgatattactgaatcacctgtcagttcattacctctcgataaggcaacagtaaacgtaaatgttc
gtgttgttgatgatgtaaaagacgaacgacagaatatttcggttgtttcaggtgttccgatgagtgttccggtggttgat
gcaaaacctaccgaacgtccaggtgtttttacggcatcaattccaggtgcacctgttctgaatatttcagttaataacag
tacgccagaagtacagacattaagcccaggtgttacaaataatactgataaggatgttcgcccggcaggatttactcagg
gtggtaataccagggatgcagttattcgattcccgaaggacagcggtcataatgccgtatatgtttcagtgagtgatgtt
cttagtcctgaccaggtaaaacaacgtcaggatgaagaaaatcgccgtcagcaggaatgggatgctacgcatccggttga
agcggctgagcgaaattatgaacgcgcgcgtgcagagctgaatcaggcaaatgaagatgttgccagaaatcaggagcgac
aggctaaagctgttcaggtttataattcgcgtaaaagcgaacttgatgcagcgaataaaactcttgctgatgcaatagct
gaaataaaacaatttaatcgatttgcccatgacccaatggctggcggtcacagaatgtggcaaatggccggacttaaagc
tcagcgggcgcagacggatgtaaataataagcaggctgcatttgatgctgctgcaaaagagaagtcagatgctgatgctg
cattaagtgccgcgcaggagcgccgcaaacagaaggaaaataaagaaaaggacgctaaggataaattagataaggagagt
aaacggaataagccagggaaggcgacaggtaaaggtaaaccagttggtgataaatggctggatgatgcaggtaaagattc
aggagcgccaattccagatcgcattgctgataagttgcgtgataaagaatttaaaaactttgacgatttccggaagaaat
tctgggaagaagtgtcaaaagatcccgatcttagtaagcaatttaaaggcagtaataagacgaacattcaaaagggaaaa
gcaccttttgcaaggaagaaagaccaagtaggtggtagggaacgctttgaattacatcatgataaaccaatcagtcagga
tggtggtgtctatgatatgaataatatcagagtgaccacacctaagcgacatattgatattcatcggggtaagtaaaaat
ggaactgaaacatagtattagtgattataccgaggctgaatttctggagtttgtaaaaaaaatatgtagagctgaaggtg
ctactgaagaggatgacaataaattagtgagagagtttgagcgattaactgagcacccagatggttcagatctgatttat
tatcctcgcgatgacagggaagatagtcctgaagggattgtcaaggaaattaaagaatggcgagctgctaacggtaagtc
aggatttaaacagggctgaaatatgaatgccggttgtttatggatgaatggctggcattctttcacaacaaggagtcgtt
atgaaaaaaataacagggattattttattgcttcttgcagtcattattctgtctgcatgtcaggcaaactatatccggga
tgttcagggcgggaccgtatctccgtcatcaacagctgaagtgaccggattagcaacgcagtaacccgaaatcctctttg
acaaaaacaaagcgtgtcaggctgattctgatgcgcttttttttgaaatgtcacaaaaattccatgtgggagatgggatc
>pBca9145-Bca1089
gaattcatgAGATCTatggcttcctccgaagacgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgt
taacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagtta
ccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacac
ccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagacgg
tggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaact
tcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagacggt
gctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacat
ggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactaca
ccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataaGGATCCtaaCTCGAGctgcaggct
tcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggtt
atccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccg
cgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaac
ccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttac
cggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgt
aggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgt
cttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgt
aggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgc
tgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggttttttt
gtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctca
gtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaa
aatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacct
atctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggctt
accatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccag
ccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctaga
gtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttgg
tatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagct
ccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattct
cttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcg
gcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattg
gaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcaccc
aactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaaggg
aataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtc
tcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgcca
cctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggcagaatttcagataaaa
aaaatccttagctttcgctaaggatgatttctg