LOCUS       pBE1203_R134E           6126 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:6xHis-TEV-MlaC(R134E); IPTG inducible "
            "Backbone:pBE1203" "Construction:T7 pro and term & MM127 & MM128
            primers used in a 3-way Gibson assembly into the NdeI and BsaI
            digested 1203 plasmid" "Resistance:Carb" "Location:Comet (-20 C)"
            "Rack" "Box" "Notes:https\://benchling.com/dcekiert/f/lib_CXPgeOQY-
            cloning/etr_y5iDHKcj-cloning-high-priority-mutants-for-expression-p
            urification/edit" "Sequencing information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pBE1203.ape
            ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    19..20
                     /label="Three base pair deletion between these two nucleotides"
     CDS             complement(1214..2293)
                     /label="lacI"
     promoter        2680..2696
                     /label="T7 promoter"
     primer_bind     2680..2699
                     /label="T7"
     misc_binding    2699..2721
                     /label="LacO"
     CDS             2699..2723
                     /label="lacO"
     misc_feature    2751..2758
                     /label="Destoryed SfiI site"
     misc_feature    2769..2771
                     /label="Start"
     CDS             2769..3380
                     /label="Translation 2769-3380"
                     /translation="MHHHHHHENLYFQADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIEVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEGVSMITTKQNEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     misc_feature    2772..2789
                     /label="His Tag"
     misc_feature    2790..2807
                     /label="TEV Cleavage Site"
     misc_feature    2808..3377
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     primer          3113..3172
                     /label="MM127"
                     /note="sequence: gccgctgggcgataaaaccattgtgcctattGAAgttaccattattgacccgaatggccg"
     primer          complement(3117..3176)
                     /label="MM128"
                     /note="sequence: cggacggccattcgggtcaataatggtaacTTCaataggcacaatggttttatcgcccag"
     misc_feature    3378..3380
                     /label="STOP"
     misc_feature    3389..3396
                     /label="Destroyed SfiI site"
     terminator      3461..3507
                     /label="T7 terminator"
     rep_origin      3544..3999
                     /label="M13 origin"
     rep_origin      3544..3999
                     /label="f1 origin"
     rep_origin      3561..3867
                     /label="F1 ori"
     misc_feature    3990..3990
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    3999..4000
                     /label="Single nucleotide deletion between these two base pairs"
     misc_feature    complement(4020..4180)
                     /label="Fragment from pL452"
     misc_recomb     4074..4107
                     /label="LoxP"
     gene            complement(4074..4107)
                     /label="loxP"
     CDS             4291..5006
                     /label="bla"
     CDS             4489..5006
                     /label="AmpR"
     CDS             5008..5148
                     /label="AmpR"
     CDS             5008..5148
                     /label="bla"
     rep_origin      5246..5928
                     /label="ColE1 origin"
ORIGIN
        1 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
       61 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
      121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      241 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
      301 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
      361 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
      421 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
      481 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
      541 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
      601 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
      661 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
      721 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
      781 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
      841 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
      901 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
      961 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     1021 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     1081 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     1141 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     1201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     1321 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     1381 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     1441 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     1501 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     1561 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     1621 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     1681 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     1741 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     1801 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     1861 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     1921 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     1981 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     2041 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     2101 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     2161 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     2221 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     2281 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     2341 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     2401 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     2461 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     2521 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     2581 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     2641 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg
     2701 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt GGCCCAGGag
     2761 atatacatAT GCATCACCAC CATCACCATg aaaatctgta ttttcaggca gaccagacca
     2821 atccgtataa gctgatggac gaggcggcgc agaaaacgtt cgatcgcctg aagaatgagc
     2881 aaccgcaaat tcgggccaac ccggattatc tgcgtaccat tgttgatcag gaactgctgc
     2941 catacgtaca ggtgaaatac gccggtgcgc tggtgctggg ccagtattac aagagtgcga
     3001 cccctgctca acgtgaagcc tactttgccg ctttccgtga gtacctgaag caggcttacg
     3061 gtcaggcgct ggcgatgtat cacggtcaaa cctatcagat tgcgccagaa cagccgctgg
     3121 gcgataaaac cattgtgcct attGAAgtta ccattattga cccgaatggc cgtccgccgg
     3181 tgcgtctgga cttccagtgg cgtaaaaact cccagacggg caattggcag gcttacgaca
     3241 tgattgctga aggcgtcagt atgatcacca ccaaacaaaa cgagtgggga acgctgctgc
     3301 gtaccaaagg tatcgacggc ctgactgcgc aactgaaatc gatttctcaa cagaaaatca
     3361 ctctggaaga gaaaaaataa gagagaccGG CCGGCCgatc cggctgctaa caaagcccga
     3421 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     3481 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     3541 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3601 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3661 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3721 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3781 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3841 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3901 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3961 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaC
     4021 GACGACCTGC AGCCAAGCTA GCTTGGCTGG ACGTAAACTC CTCTTCAGAC CTAATAACTT
     4081 CGTATAGCAT ACATTATACG AAGTTATATT AAGGGTTATT GAATATGATC GGAATTGGGC
     4141 TGCAGGAATT CGATAGCTTG GCTGCAGGTC GACCTCGAGG atgtgcgcgg aacccctatt
     4201 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4261 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4321 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4381 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4441 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4501 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4561 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4621 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4681 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4741 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4801 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     4861 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4921 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4981 gataaatctg gagccggtga gcgtggctct cgcggtatca ttgcagcact ggggccagat
     5041 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5101 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5161 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5221 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5281 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5341 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5401 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5461 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5521 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5581 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5641 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5701 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5761 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5821 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5881 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5941 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6001 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6061 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     6121 catctg
//