LOCUS       pBE1160_mutL123A        6012 bp ds-DNA     circular     14-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:Contains MlaD with Leucine mutated to Alanine at
            original bp 123 (105 on this one)" "Backbone:pBE1092"
            "Construction:pBE1092 digest B Fragment 1\: oLK005 + oET002
            Fragment 2\: oLK006 + oET001" "Resistance:Carb" "Location:Comet
            (-20 C)" "Rack:3" "Box:#2-Plasmids" "Notes" "Sequencing
            information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pBE1160 (1).ape
            ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    70..71
                     /label="Three base pair deletion between these two base pairs"
     CDS             complement(1265..2344)
                     /label="lacI"
     promoter        2731..2747
                     /label="T7 promoter"
     primer_bind     2731..2750
                     /label="T7"
     misc_binding    2750..2772
                     /label="LacO"
     CDS             2750..2774
                     /label="lacO"
     misc_feature    2802..2809
                     /label="Destoryed SfiI site"
     misc_feature    2820..2822
                     /label="Start"
     CDS             2820..3317
                     /label="Translation 2820-3317"
                     /translation="MHHHHHHENLYFQTSIRTEPTYTLYATFDNIGGLKARSPVSIGGVVVGRVADITLDPKTYLPRVTLEIEQRYNHIPDTSSLSIRTSGLLGEQYLALNVGFEDPEAGTAILKDGDTIQDTKSAMVLEDLIGQFLYGSKGDDNKNSGDAPAAAPGNNETTEPVGTTK*"
     misc_feature    2823..2840
                     /label="His Tag"
     primer          2826..2882
                     /label="FwdpBE1326mut"
                     /note="sequence: CACCACCATCACCATgaaaatctgtattttcagacgtccatacgtactgaaccgacc"
     misc_feature    2841..2858
                     /label="TEV Cleavage Site"
     CDS             2859..3314
                     /label="OM lipid asymmetry maintenance protein; membrane-a"
     misc_feature    2859..3314
                     /label="mlaD"
     misc_feature    2877..3116
                     /label="MCE Domain (CDD)"
     primer          3109..3158
                     /label="FwdmutL123A"
                     /note="sequence: acgtcggttttgaagacccggaaGCGgggactgctatcctgaaggatggc"
     primer          complement(3109..3158)
                     /label="RevmutL123A"
                     /note="sequence: gccatccttcaggatagcagtcccCGCttccgggtcttcaaaaccgacgt"
     misc_feature    3132..3134
                     /label="mutL_A"
     primer          complement(3289..3333)
                     /label="RevpBE1326mut"
                     /note="sequence: GGCCGGCCggtctctcttatttcgttgtacccacaggttcagtgg"
     misc_feature    3315..3317
                     /label="STOP"
     misc_feature    3326..3333
                     /label="Destroyed SfiI site"
     terminator      3398..3444
                     /label="T7 terminator"
     rep_origin      3481..3936
                     /label="M13 origin"
     rep_origin      3481..3936
                     /label="f1 origin"
     rep_origin      3498..3804
                     /label="F1 ori"
     misc_feature    3927..3927
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    3936..3937
                     /label="Single nucleotide deletion between these two base pairs"
     misc_feature    complement(3957..4117)
                     /label="Fragment from pL452"
     misc_recomb     4011..4044
                     /label="LoxP"
     gene            complement(4011..4044)
                     /label="loxP"
     CDS             4228..4943
                     /label="bla"
     CDS             4426..4943
                     /label="AmpR"
     CDS             4945..5085
                     /label="AmpR"
     CDS             4945..5085
                     /label="bla"
     rep_origin      5183..5865
                     /label="ColE1 origin"
ORIGIN
        1 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
       61 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      121 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
      181 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      241 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
      301 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
      361 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
      421 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
      481 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
      541 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
      601 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
      661 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
      721 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
      781 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
      841 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
      901 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
      961 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     1021 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     1081 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     1141 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     1201 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     1261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1321 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     1381 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     1441 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     1501 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     1561 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     1621 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     1681 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     1741 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     1801 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     1861 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     1921 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     1981 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     2041 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     2101 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     2161 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     2221 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     2281 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     2341 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     2401 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     2461 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     2521 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     2581 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     2641 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     2701 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     2761 cggataacaa ttcccctcta gaaataattt tgtttaactt tGGCCCAGGa gatatacatA
     2821 TGCATCACCA CCATCACCAT gaaaatctgt attttcagac gtccatacgt actgaaccga
     2881 cctacacgct ttatgcgacg ttcgataaca ttggcggcct gaaagcgcgc tctccggtca
     2941 gtattggtgg cgttgttgtg ggtcgggtgg cggatattac gctggacccg aaaacctatc
     3001 tgccgcgcgt aacgctggaa attgaacaac gttataacca cattccagat accagttcgc
     3061 tgagcattcg tacttccggc ctgctggggg aacaatatct ggcattaaac gtcggttttg
     3121 aagacccgga aGCGgggact gctatcctga aggatggcga tacaattcag gacactaagt
     3181 ctgcgatggt gctggaagat ctcattggtc agttccttta cggtagtaaa ggcgatgaca
     3241 ataagaatag tggcgatgcg ccagctgctg cgccaggtaa taatgaaacc actgaacctg
     3301 tgggtacaac gaaataagag agaccGGCCG GCCgatccgg ctgctaacaa agcccgaaag
     3361 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     3421 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     3481 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3541 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3601 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3661 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3721 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3781 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3841 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3901 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaCGAC
     3961 GACCTGCAGC CAAGCTAGCT TGGCTGGACG TAAACTCCTC TTCAGACCTA ATAACTTCGT
     4021 ATAGCATACA TTATACGAAG TTATATTAAG GGTTATTGAA TATGATCGGA ATTGGGCTGC
     4081 AGGAATTCGA TAGCTTGGCT GCAGGTCGAC CTCGAGGatg tgcgcggaac ccctatttgt
     4141 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4201 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4261 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4321 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4381 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4441 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4501 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4561 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4621 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4681 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4741 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4801 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4861 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4921 aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg gccagatggt
     4981 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5041 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5101 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5161 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5401 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6001 agcgagtcag tg
//