LOCUS       pBE1203_D165I           6126 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:6xHis-TEV-MlaC(D165I); IPTG inducible "
            "Backbone:pBE1203" "Construction:T7 pro and term & MM112 and MM111
            primers used in a 3-way gibson assembly into the NdeI and BsaI
            digested pBEL1203 plasmid" "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
     CDS             complement(1125..2204)
                     /label="lacI"
     promoter        2591..2607
                     /label="T7 promoter"
     primer_bind     2591..2610
                     /label="T7"
     misc_binding    2610..2632
                     /label="LacO"
     CDS             2610..2634
                     /label="lacO"
     misc_feature    2662..2669
                     /label="Destoryed SfiI site"
     misc_feature    2680..2682
                     /label="Start"
     CDS             2680..3291
                     /label="Translation 2680-3291"
                     /translation="MHHHHHHENLYFQADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYIMIAEGVSMITTKQNEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     misc_feature    2683..2700
                     /label="His Tag"
     misc_feature    2701..2718
                     /label="TEV Cleavage Site"
     misc_feature    2719..3288
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     primer          3119..3178
                     /label="MM111"
                     /note="sequence: actcccagacgggcaattggcaggcttacATTatgattgctgaaggcgtcagtatgatca"
     primer          complement(3127..3186)
                     /label="MM112"
                     /note="sequence: tttggtggtgatcatactgacgccttcagcaatcatAATgtaagcctgccaattgcccgt"
     misc_feature    3289..3291
                     /label="STOP"
     misc_feature    3300..3307
                     /label="Destroyed SfiI site"
     terminator      3372..3418
                     /label="T7 terminator"
     rep_origin      3455..3910
                     /label="M13 origin"
     rep_origin      3455..3910
                     /label="f1 origin"
     rep_origin      3472..3778
                     /label="F1 ori"
     misc_feature    3901..3901
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    3910..3911
                     /label="Single nucleotide deletion between these two base pairs"
     misc_feature    complement(3931..4091)
                     /label="Fragment from pL452"
     misc_recomb     3985..4018
                     /label="LoxP"
     gene            complement(3985..4018)
                     /label="loxP"
     CDS             4202..4917
                     /label="bla"
     CDS             4400..4917
                     /label="AmpR"
     CDS             4919..5059
                     /label="AmpR"
     CDS             4919..5059
                     /label="bla"
     rep_origin      5157..5839
                     /label="ColE1 origin"
     misc_feature    6056..6057
                     /label="Three base pair deletion between these two nucleotides"
ORIGIN
        1 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
       61 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
      181 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
      241 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
      301 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
      361 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
      421 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
      481 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
      541 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
      601 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
      661 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
      721 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
      781 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
      841 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
      901 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
      961 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     1021 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     1081 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     1141 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1201 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     1261 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     1321 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     1381 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     1441 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     1501 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     1561 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     1621 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     1681 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     1741 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     1801 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     1861 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     1921 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     1981 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     2041 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     2101 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     2161 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     2221 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     2281 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     2341 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     2401 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     2461 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     2521 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     2581 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2641 gaaataattt tgtttaactt tGGCCCAGGa gatatacatA TGCATCACCA CCATCACCAT
     2701 gaaaatctgt attttcaggc agaccagacc aatccgtata agctgatgga cgaggcggcg
     2761 cagaaaacgt tcgatcgcct gaagaatgag caaccgcaaa ttcgggccaa cccggattat
     2821 ctgcgtacca ttgttgatca ggaactgctg ccatacgtac aggtgaaata cgccggtgcg
     2881 ctggtgctgg gccagtatta caagagtgcg acccctgctc aacgtgaagc ctactttgcc
     2941 gctttccgtg agtacctgaa gcaggcttac ggtcaggcgc tggcgatgta tcacggtcaa
     3001 acctatcaga ttgcgccaga acagccgctg ggcgataaaa ccattgtgcc tattcgcgtt
     3061 accattattg acccgaatgg ccgtccgccg gtgcgtctgg acttccagtg gcgtaaaaac
     3121 tcccagacgg gcaattggca ggcttacATT atgattgctg aaggcgtcag tatgatcacc
     3181 accaaacaaa acgagtgggg aacgctgctg cgtaccaaag gtatcgacgg cctgactgcg
     3241 caactgaaat cgatttctca acagaaaatc actctggaag agaaaaaata agagagaccG
     3301 GCCGGCCgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     3361 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     3421 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     3481 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3541 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3601 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3661 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3721 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3781 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3841 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3901 cttacaattt aggtggcact tttcggggaa CGACGACCTG CAGCCAAGCT AGCTTGGCTG
     3961 GACGTAAACT CCTCTTCAGA CCTAATAACT TCGTATAGCA TACATTATAC GAAGTTATAT
     4021 TAAGGGTTAT TGAATATGAT CGGAATTGGG CTGCAGGAAT TCGATAGCTT GGCTGCAGGT
     4081 CGACCTCGAG Gatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4141 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4201 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4261 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4321 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4381 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4441 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4501 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4561 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4621 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4681 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4741 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4801 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4861 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggctc
     4921 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4981 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5041 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5341 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5881 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5941 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6001 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     6061 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     6121 cgctac
//