LOCUS       pBE1203_L76D            6126 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:6xHis-TEV-MlaC(L76D); IPTG inducible "
            "Backbone:pBE1203" "Construction:T7 pro and term & MM122 and MM121
            primers used in 3-way gibson assembly 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
     rep_origin      216..898
                     /label="ColE1 origin"
     misc_feature    1115..1116
                     /label="Three base pair deletion between these two nucleotides"
     CDS             complement(2310..3389)
                     /label="lacI"
     promoter        3776..3792
                     /label="T7 promoter"
     primer_bind     3776..3795
                     /label="T7"
     misc_binding    3795..3817
                     /label="LacO"
     CDS             3795..3819
                     /label="lacO"
     misc_feature    3847..3854
                     /label="Destoryed SfiI site"
     misc_feature    3865..3867
                     /label="Start"
     CDS             3865..4476
                     /label="Translation 3865-4476"
                     /translation="MHHHHHHENLYFQADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGADVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEGVSMITTKQNEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     misc_feature    3868..3885
                     /label="His Tag"
     misc_feature    3886..3903
                     /label="TEV Cleavage Site"
     misc_feature    3904..4473
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     primer          4034..4093
                     /label="MM121"
                     /note="sequence: tgccatacgtacaggtgaaatacgccggtgcgGATgtgctgggccagtattacaagagtg"
     primer          complement(4046..4105)
                     /label="MM122"
                     /note="sequence: gagcaggggtcgcactcttgtaatactggcccagcacATCcgcaccggcgtatttcacct"
     misc_feature    4474..4476
                     /label="STOP"
     misc_feature    4485..4492
                     /label="Destroyed SfiI site"
     terminator      4557..4603
                     /label="T7 terminator"
     rep_origin      4640..5095
                     /label="f1 origin"
     rep_origin      4640..5095
                     /label="M13 origin"
     rep_origin      4657..4963
                     /label="F1 ori"
     misc_feature    5086..5086
                     /label="Point mutation: Thymine to cytosine"
     misc_feature    5095..5096
                     /label="One nucleotide deletion between these two base pairs"
     misc_feature    complement(5116..5276)
                     /label="Fragment from pL452"
     misc_recomb     5170..5203
                     /label="LoxP"
     gene            complement(5170..5203)
                     /label="loxP"
     CDS             5387..6102
                     /label="bla"
     CDS             5585..6102
                     /label="AmpR"
     CDS             6104..118
                     /label="AmpR"
     CDS             6104..118
                     /label="bla"
ORIGIN
        1 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
       61 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
      121 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
      181 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
      241 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
      301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      421 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      781 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      901 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      961 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1021 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     1081 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     1141 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     1201 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     1261 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     1321 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     1381 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     1441 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     1501 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     1561 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     1621 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     1681 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     1741 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     1801 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     1861 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     1921 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     1981 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     2041 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     2101 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     2161 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     2221 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     2281 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2341 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     2401 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     2461 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     2521 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     2581 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     2641 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     2701 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     2761 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     2821 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     2881 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     2941 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     3001 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     3061 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     3121 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     3181 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     3241 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     3301 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     3361 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     3421 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     3481 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     3541 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     3601 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     3661 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     3721 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
     3781 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt
     3841 aactttGGCC CAGGagatat acatATGCAT CACCACCATC ACCATgaaaa tctgtatttt
     3901 caggcagacc agaccaatcc gtataagctg atggacgagg cggcgcagaa aacgttcgat
     3961 cgcctgaaga atgagcaacc gcaaattcgg gccaacccgg attatctgcg taccattgtt
     4021 gatcaggaac tgctgccata cgtacaggtg aaatacgccg gtgcgGATgt gctgggccag
     4081 tattacaaga gtgcgacccc tgctcaacgt gaagcctact ttgccgcttt ccgtgagtac
     4141 ctgaagcagg cttacggtca ggcgctggcg atgtatcacg gtcaaaccta tcagattgcg
     4201 ccagaacagc cgctgggcga taaaaccatt gtgcctattc gcgttaccat tattgacccg
     4261 aatggccgtc cgccggtgcg tctggacttc cagtggcgta aaaactccca gacgggcaat
     4321 tggcaggctt acgacatgat tgctgaaggc gtcagtatga tcaccaccaa acaaaacgag
     4381 tggggaacgc tgctgcgtac caaaggtatc gacggcctga ctgcgcaact gaaatcgatt
     4441 tctcaacaga aaatcactct ggaagagaaa aaataagaga gaccGGCCGG CCgatccggc
     4501 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     4561 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     4621 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4681 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4741 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4801 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4861 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4921 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4981 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5041 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     5101 gcacttttcg gggaaCGACG ACCTGCAGCC AAGCTAGCTT GGCTGGACGT AAACTCCTCT
     5161 TCAGACCTAA TAACTTCGTA TAGCATACAT TATACGAAGT TATATTAAGG GTTATTGAAT
     5221 ATGATCGGAA TTGGGCTGCA GGAATTCGAT AGCTTGGCTG CAGGTCGACC TCGAGGatgt
     5281 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5341 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5401 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5461 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5521 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5581 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5641 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5701 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5761 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5821 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5881 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     5941 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6001 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6061 tggctggttt attgctgata aatctggagc cggtgagcgt ggctctcgcg gtatcattgc
     6121 agcact
//