LOCUS       pBE1203_N179E           6126 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:6xHis-TEV-MlaC(N179E); IPTG inducible "
            "Backbone:pBE1203" "Construction:T7 pro and term & MM157 & MM158
            primers used in 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
     CDS             complement(1174..2253)
                     /label="lacI"
     promoter        2640..2656
                     /label="T7 promoter"
     primer_bind     2640..2659
                     /label="T7"
     misc_binding    2659..2681
                     /label="LacO"
     CDS             2659..2683
                     /label="lacO"
     misc_feature    2711..2718
                     /label="Destoryed SfiI site"
     misc_feature    2729..2731
                     /label="Start"
     CDS             2729..3340
                     /label="Translation 2729-3340"
                     /translation="MHHHHHHENLYFQADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEGVSMITTKQEEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     misc_feature    2732..2749
                     /label="His Tag"
     misc_feature    2750..2767
                     /label="TEV Cleavage Site"
     misc_feature    2768..3337
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     primer          3201..3260
                     /label="MM157"
                     /note="sequence: tgattgctgaaggcgtcagtatgatcaccaccaaacaaGAAgagtggggaacgctgctgc"
     primer          complement(3213..3272)
                     /label="MM158"
                     /note="sequence: tacctttggtacgcagcagcgttccccactcTTCttgtttggtggtgatcatactgacgc"
     misc_feature    3338..3340
                     /label="STOP"
     misc_feature    3349..3356
                     /label="Destroyed SfiI site"
     terminator      3421..3467
                     /label="T7 terminator"
     rep_origin      3504..3959
                     /label="f1 origin"
     rep_origin      3504..3959
                     /label="M13 origin"
     rep_origin      3521..3827
                     /label="F1 ori"
     misc_feature    3950..3950
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    3959..3960
                     /label="Single nucleotide deletion between these two base pairs"
     misc_feature    complement(3980..4140)
                     /label="Fragment from pL452"
     misc_recomb     4034..4067
                     /label="LoxP"
     gene            complement(4034..4067)
                     /label="loxP"
     CDS             4251..4966
                     /label="bla"
     CDS             4449..4966
                     /label="AmpR"
     CDS             4968..5108
                     /label="bla"
     CDS             4968..5108
                     /label="AmpR"
     rep_origin      5206..5888
                     /label="ColE1 origin"
     misc_feature    6105..6106
                     /label="Three base pair deletion between these two nucleotides"
ORIGIN
        1 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
       61 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
      121 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
      181 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
      241 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
      301 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
      361 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
      421 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
      481 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
      541 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
      601 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
      661 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
      721 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
      781 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
      841 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
      901 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
      961 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     1021 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     1081 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     1141 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1201 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1261 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     1321 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     1381 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     1441 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     1501 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     1561 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     1621 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     1681 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     1741 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     1801 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     1861 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     1921 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     1981 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     2041 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     2101 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     2161 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     2221 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     2281 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     2341 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     2401 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     2461 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     2521 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     2581 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     2641 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     2701 gtttaacttt GGCCCAGGag atatacatAT GCATCACCAC CATCACCATg aaaatctgta
     2761 ttttcaggca gaccagacca atccgtataa gctgatggac gaggcggcgc agaaaacgtt
     2821 cgatcgcctg aagaatgagc aaccgcaaat tcgggccaac ccggattatc tgcgtaccat
     2881 tgttgatcag gaactgctgc catacgtaca ggtgaaatac gccggtgcgc tggtgctggg
     2941 ccagtattac aagagtgcga cccctgctca acgtgaagcc tactttgccg ctttccgtga
     3001 gtacctgaag caggcttacg gtcaggcgct ggcgatgtat cacggtcaaa cctatcagat
     3061 tgcgccagaa cagccgctgg gcgataaaac cattgtgcct attcgcgtta ccattattga
     3121 cccgaatggc cgtccgccgg tgcgtctgga cttccagtgg cgtaaaaact cccagacggg
     3181 caattggcag gcttacgaca tgattgctga aggcgtcagt atgatcacca ccaaacaaGA
     3241 Agagtgggga acgctgctgc gtaccaaagg tatcgacggc ctgactgcgc aactgaaatc
     3301 gatttctcaa cagaaaatca ctctggaaga gaaaaaataa gagagaccGG CCGGCCgatc
     3361 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     3421 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     3481 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3841 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3901 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3961 ggtggcactt ttcggggaaC GACGACCTGC AGCCAAGCTA GCTTGGCTGG ACGTAAACTC
     4021 CTCTTCAGAC CTAATAACTT CGTATAGCAT ACATTATACG AAGTTATATT AAGGGTTATT
     4081 GAATATGATC GGAATTGGGC TGCAGGAATT CGATAGCTTG GCTGCAGGTC GACCTCGAGG
     4141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4201 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4261 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4321 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4381 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4441 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4501 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4561 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4621 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4681 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4741 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     4801 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4861 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4921 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggctct cgcggtatca
     4981 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5041 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5101 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5161 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5221 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5281 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5341 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5401 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5461 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5521 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5581 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5641 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5701 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5761 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5821 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5881 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5941 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6001 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     6061 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     6121 tacaat
//