LOCUS       pBE1203_G170K           6126 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:6xHis-TEV-MlaC(G170K); IPTG inducible "
            "Backbone:pBE1203" "Construction:T7 pro and term & MM155 & MM156
            used in 3-way gibson assembly into the NdeI and BsaI digested
            pBEL1203 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-purifica
            tion/edit" "Sequencing information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pBE1203.ape
            ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     CDS             complement(1032..2111)
                     /label="lacI"
     promoter        2498..2514
                     /label="T7 promoter"
     primer_bind     2498..2517
                     /label="T7"
     misc_binding    2517..2539
                     /label="LacO"
     CDS             2517..2541
                     /label="lacO"
     misc_feature    2569..2576
                     /label="Destoryed SfiI site"
     misc_feature    2587..2589
                     /label="Start"
     CDS             2587..3198
                     /label="Translation 2587-3198"
                     /translation="MHHHHHHENLYFQADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEKVSMITTKQNEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     misc_feature    2590..2607
                     /label="His Tag"
     misc_feature    2608..2625
                     /label="TEV Cleavage Site"
     misc_feature    2626..3195
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     primer          3042..3101
                     /label="MM155"
                     /note="sequence: ttggcaggcttacgacatgattgctgaaAAAgtcagtatgatcaccaccaaacaaaacga"
     primer          complement(3045..3104)
                     /label="MM156"
                     /note="sequence: cactcgttttgtttggtggtgatcatactgacTTTttcagcaatcatgtcgtaagcctgc"
     misc_feature    3196..3198
                     /label="STOP"
     misc_feature    3207..3214
                     /label="Destroyed SfiI site"
     terminator      3279..3325
                     /label="T7 terminator"
     rep_origin      3362..3817
                     /label="M13 origin"
     rep_origin      3362..3817
                     /label="f1 origin"
     rep_origin      3379..3685
                     /label="F1 ori"
     misc_feature    3808..3808
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    3817..3818
                     /label="One nucleotide deletion between these two base pairs"
     misc_feature    complement(3838..3998)
                     /label="Fragment from pL452"
     misc_recomb     3892..3925
                     /label="LoxP"
     gene            complement(3892..3925)
                     /label="loxP"
     CDS             4109..4824
                     /label="bla"
     CDS             4307..4824
                     /label="AmpR"
     CDS             4826..4966
                     /label="bla"
     CDS             4826..4966
                     /label="AmpR"
     rep_origin      5064..5746
                     /label="ColE1 origin"
     misc_feature    5963..5964
                     /label="Three base pair deletion between these two nucleotides"
ORIGIN
        1 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
       61 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
      121 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
      181 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
      241 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
      301 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
      361 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
      421 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
      481 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
      541 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
      601 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
      661 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
      721 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
      781 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
      841 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
      901 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
      961 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     1021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     1141 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     1201 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     1261 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     1321 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     1381 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     1441 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     1501 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     1561 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     1621 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     1681 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     1741 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     1801 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     1861 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     1921 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     1981 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     2041 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     2101 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     2161 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     2221 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     2281 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     2341 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     2401 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     2461 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     2521 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttGG CCCAGGagat
     2581 atacatATGC ATCACCACCA TCACCATgaa aatctgtatt ttcaggcaga ccagaccaat
     2641 ccgtataagc tgatggacga ggcggcgcag aaaacgttcg atcgcctgaa gaatgagcaa
     2701 ccgcaaattc gggccaaccc ggattatctg cgtaccattg ttgatcagga actgctgcca
     2761 tacgtacagg tgaaatacgc cggtgcgctg gtgctgggcc agtattacaa gagtgcgacc
     2821 cctgctcaac gtgaagccta ctttgccgct ttccgtgagt acctgaagca ggcttacggt
     2881 caggcgctgg cgatgtatca cggtcaaacc tatcagattg cgccagaaca gccgctgggc
     2941 gataaaacca ttgtgcctat tcgcgttacc attattgacc cgaatggccg tccgccggtg
     3001 cgtctggact tccagtggcg taaaaactcc cagacgggca attggcaggc ttacgacatg
     3061 attgctgaaA AAgtcagtat gatcaccacc aaacaaaacg agtggggaac gctgctgcgt
     3121 accaaaggta tcgacggcct gactgcgcaa ctgaaatcga tttctcaaca gaaaatcact
     3181 ctggaagaga aaaaataaga gagaccGGCC GGCCgatccg gctgctaaca aagcccgaaa
     3241 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     3301 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     3361 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3421 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3481 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3541 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3601 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3661 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3721 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3781 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaCGA
     3841 CGACCTGCAG CCAAGCTAGC TTGGCTGGAC GTAAACTCCT CTTCAGACCT AATAACTTCG
     3901 TATAGCATAC ATTATACGAA GTTATATTAA GGGTTATTGA ATATGATCGG AATTGGGCTG
     3961 CAGGAATTCG ATAGCTTGGC TGCAGGTCGA CCTCGAGGat gtgcgcggaa cccctatttg
     4021 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4081 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4141 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4201 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4261 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4321 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4381 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4441 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4501 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4561 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4621 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     4681 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4741 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4801 taaatctgga gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg
     4861 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4921 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4981 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5041 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5281 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5401 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5461 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5521 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5581 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5641 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5701 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5761 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5821 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5881 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     5941 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     6001 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     6061 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     6121 gacaag
//