LOCUS       pBE1203_T176R           6126 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:6xHis-TEV-MlaC(T176R); IPTG inducible "
            "Backbone:pBE1203" "Construction:Used T7 pro and term and MM138 and
            MM137 for 3-way gibson into NdeI and BsaI digested pBEL1203
            backbone" "Resistance:Carb" "Location:Comet (-20 C)" "Rack" "Box"
            "Notes" "Sequencing information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pBE1203.ape
            ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    1..1
                     /label="Three base pair deletion right before this nucleotide"
     CDS             complement(1195..2274)
                     /label="lacI"
     promoter        2661..2677
                     /label="T7 promoter"
     primer_bind     2661..2680
                     /label="T7"
     misc_binding    2680..2702
                     /label="LacO"
     CDS             2680..2704
                     /label="lacO"
     misc_feature    2732..2739
                     /label="Destoryed SfiI site"
     misc_feature    2750..2752
                     /label="Start"
     CDS             2750..3361
                     /label="Translation 2750-3361"
                     /translation="MHHHHHHENLYFQADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEGVSMITRKQNEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     misc_feature    2753..2770
                     /label="His Tag"
     misc_feature    2771..2788
                     /label="TEV Cleavage Site"
     misc_feature    2789..3358
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     primer          3218..3277
                     /label="MM137"
                     /note="sequence: gacatgattgctgaaggcgtcagtatgatcaccCGCaaacaaaacgagtggggaacgctg"
     primer          complement(3226..3285)
                     /label="MM138"
                     /note="sequence: gtacgcagcagcgttccccactcgttttgtttGCGggtgatcatactgacgccttcagca"
     misc_feature    3359..3361
                     /label="STOP"
     misc_feature    3370..3377
                     /label="Destroyed SfiI site"
     terminator      3442..3488
                     /label="T7 terminator"
     rep_origin      3525..3980
                     /label="f1 origin"
     rep_origin      3525..3980
                     /label="M13 origin"
     rep_origin      3542..3848
                     /label="F1 ori"
     misc_feature    3971..3971
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    3980..3981
                     /label="Single nucleotide deletion between these two base pairs"
     misc_feature    complement(4001..4161)
                     /label="Fragment from pL452"
     misc_recomb     4055..4088
                     /label="LoxP"
     gene            complement(4055..4088)
                     /label="loxP"
     CDS             4272..4987
                     /label="bla"
     CDS             4470..4987
                     /label="AmpR"
     CDS             4989..5129
                     /label="bla"
     CDS             4989..5129
                     /label="AmpR"
     rep_origin      5227..5909
                     /label="ColE1 origin"
ORIGIN
        1 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
       61 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
      121 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
      181 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
      241 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
      301 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
      361 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
      421 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
      481 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
      541 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
      601 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
      661 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
      721 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
      781 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
      841 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
      901 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
      961 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1021 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1081 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     1141 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     1201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1261 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     1321 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     1381 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     1441 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     1501 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     1561 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     1621 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     1681 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     1741 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     1801 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     1861 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     1921 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     1981 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2041 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     2101 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     2161 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     2221 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     2281 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     2341 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     2401 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     2461 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     2521 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     2581 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     2641 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     2701 ttcccctcta gaaataattt tgtttaactt tGGCCCAGGa gatatacatA TGCATCACCA
     2761 CCATCACCAT gaaaatctgt attttcaggc agaccagacc aatccgtata agctgatgga
     2821 cgaggcggcg cagaaaacgt tcgatcgcct gaagaatgag caaccgcaaa ttcgggccaa
     2881 cccggattat ctgcgtacca ttgttgatca ggaactgctg ccatacgtac aggtgaaata
     2941 cgccggtgcg ctggtgctgg gccagtatta caagagtgcg acccctgctc aacgtgaagc
     3001 ctactttgcc gctttccgtg agtacctgaa gcaggcttac ggtcaggcgc tggcgatgta
     3061 tcacggtcaa acctatcaga ttgcgccaga acagccgctg ggcgataaaa ccattgtgcc
     3121 tattcgcgtt accattattg acccgaatgg ccgtccgccg gtgcgtctgg acttccagtg
     3181 gcgtaaaaac tcccagacgg gcaattggca ggcttacgac atgattgctg aaggcgtcag
     3241 tatgatcacc CGCaaacaaa acgagtgggg aacgctgctg cgtaccaaag gtatcgacgg
     3301 cctgactgcg caactgaaat cgatttctca acagaaaatc actctggaag agaaaaaata
     3361 agagagaccG GCCGGCCgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3421 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     3481 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     3541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3601 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3721 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3841 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3961 aatattaacg cttacaattt aggtggcact tttcggggaa CGACGACCTG CAGCCAAGCT
     4021 AGCTTGGCTG GACGTAAACT CCTCTTCAGA CCTAATAACT TCGTATAGCA TACATTATAC
     4081 GAAGTTATAT TAAGGGTTAT TGAATATGAT CGGAATTGGG CTGCAGGAAT TCGATAGCTT
     4141 GGCTGCAGGT CGACCTCGAG Gatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4381 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4441 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4501 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4561 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4621 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4681 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4741 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4801 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4861 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4921 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4981 agcgtggctc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5041 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5101 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6001 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6061 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6121 accgca
//