LOCUS       pBEL2766                7545 bp ds-DNA     circular     25-NOV-2022
DEFINITION  .
KEYWORDS    "Key Features:mlaFEDCB operon, MlaD(1-140)" "Backbone:pBEL1195"
            "Construction:Three-way Gibson cloning of PCR2227 and PCR2228 into
            SalI-SacII-digested pBE1195" "Resistance:Carb" "Location" "Rack"
            "Box" "Notes:REPLACES pBEL1330 from the lab collection, which was a
            dimer." "Sequencing information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pBE1330.ape
            ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     promoter        13..285
                     /label="araBAD promoter region"
     CDS             328..1137
                     /label="mlaF, ABC type ATPase"
     primer          806..827
                     /label="MM028"
                     /note="sequence: cgctggagccggatctcatcat"
     primer          922..943
                     /label="oDCE3363"
                     /note="sequence: gtggtggtttctcacgatgtgc"
     misc_feature    1145..1927
                     /label="mlaE, 5 predicted TM, possible pore"
     CDS             1145..1927
                     /translation="MLLNALASLGHKGIKTLRTFGRAGLMLFNALVGKPEFRKHAPLLVRQLYNVGVLSMLIIVVSGVFIGMVLGLQGYLVLTTYSAETSLGMLVALSLLRELGPVVAALLFAGRAGSALTAEIGLMRATEQLSSMEMMAVDPLRRVISPRFWAGVISLPLLTVIFVAVGIWGGSLVGVSWKGIDSGFFWSAMQNAVDWRMDLVNCLIKSVVFAITVTWISLFNGYDAIPTSAGISRATTRTVVHSSLAVLGLDFVLTALMFGN*"
     primer          1492..1513
                     /label="MM029"
                     /note="sequence: aaccgcagaaatcggcctgatg"
     misc_feature    1932..2354
                     /label="mlaD"
     CDS             1932..2354
                     /label="Translation 1932-2354"
                     /translation="MQTKKNEIWVGIFLLAALLAALFVCLKAANVTSIRTEPTYTLYATFDNIGGLKARSPVSIGGVVVGRVADITLDPKTYLPRVTLEIEQRYNHIPDTSSLSIRTSGLLGEQYLALNVGFEDPELGTAILKDGDTIQDTKSA*"
     misc_feature    1959..2021
                     /label="TM helix (TOPCONS)"
     misc_feature    2043..2282
                     /label="MCE Domain (CDD)"
     primer          2124..2145
                     /label="MM030"
                     /note="sequence: gtgggtcgggtggcggatatta"
     primer          2324..2383
                     /label="MM160"
                     /note="sequence: cgatacaattcaggacactaagtctgcgtaatttcaggagaaccgacgcatgtttaaacg"
     primer          complement(2324..2383)
                     /label="MM159"
                     /note="sequence: cgtttaaacatgcgtcggttctcctgaaattacgcagacttagtgtcctgaattgtatcg"
     variation       2351..2352
                     /label="Deletion residues 141-183"
     misc_feature    2373..3008
                     /label="mlaC, predicted signal peptide, possible periplasmi"
     CDS             2373..3008
                     /label="Translation 2373-3008"
                     /translation="MFKRLMMVALLVIAPLSAATAADQTNPYKLMDEAAQKTFDRLKNEQPQIRANPDYLRTIVDQELLPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIAPEQPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEGVSMITTKQNEWGTLLRTKGIDGLTAQLKSISQQKITLEEKK*"
     primer          complement(2556..2577)
                     /label="oDCE3366"
                     /note="sequence: gtacgtatggcagcagttcctg"
     CDS             3008..3301
                     /label="ABC transporter maintaining OM lipid asymmetry, cy"
     misc_feature    3008..3301
                     /label="mlaB"
     terminator      3463..3620
                     /label="rrnB transcription terminator"
     misc_feature    3599..3623
                     /label="BE2467"
     CDS             3899..4759
                     /label="Ampicillin ORF"
     misc_feature    4443..4443
                     /label="SeqResult T->C"
     rep_origin      5291..5972
                     /label="ColE1 origin"
     misc_feature    6347..6347
                     /label="SeqResC->A"
     misc_feature    6350..6352
                     /label="SeqResATC->GAG"
     misc_feature    6354..6558
                     /label="SeqRes 205bp insert"
     CDS             complement(6650..7537)
                     /label="araC"
ORIGIN
        1 CATTCAGAGA AGAAACCAAT TGTCCATATT GCATCAGACA TTGCCGTCAC TGCGTCTTTT
       61 ACTGGCTCTT CTCGCTAACC AAACCGGTAA CCCCGCTTAT TAAAAGCATT CTGTAACAAA
      121 GCGGGACCAA AGCCATGACA AAAACGCGTA ACAAAAGTGT CTATAATCAC GGCAGAAAAG
      181 TCCACATTGA TTATTTGCAC GGCGTCACAC TTTGCTATGC CATAGCATTT TTATCCATAA
      241 GATTAGCGGA TCCTACCTGA CGCTTTTTAT CGCAACTCTC TACTGTTTCT CCATACCCGT
      301 TTTTTTGGGC TAGCcgaagg gtgaattatg gagcagtctg tggcgaattt agtcgatatg
      361 cgcgatgtca gttttacgcg tggcaatcgc tgcatcttcg ataatatttc cctgaccgtg
      421 ccgcgaggga agatcacggc gatcatgggg ccatcgggca tcggtaaaac gacgctactc
      481 cgtctgattg gcgggcaaat cgcaccagat catggtgaga tccttttcga tggtgagaat
      541 attccggcga tgtctcgttc gcgcctgtat acagtgcgca aacggatgag catgttattt
      601 cagtccgggg cgttgttcac tgatatgaac gtatttgaca acgtcgccta tccactgcgc
      661 gaacataccc aacttcccgc gccattgttg catagtacgg tgatgatgaa gctggaggcc
      721 gtggggctgc gtggagcggc taaactaatg ccttctgaac tttccggtgg gatggcgcgg
      781 cgtgcagcgc tggcgcgtgc gattgcgctg gagccggatc tcatcatgtt tgatgaacct
      841 tttgttgggc aagatcccat caccatgggc gtactggtga agctgatttc tgagctgaac
      901 agcgcgctgg gcgtgacttg tgtggtggtt tctcacgatg tgccggaagt gttaagtatt
      961 gcggatcacg cctggatcct ggcggacaaa aaaattgtcg ctcatggcag tgcccaggcg
     1021 ttgcaggcga atcctgatcc gcgcgtacgt cagtttctgg acgggatagc tgacgggcct
     1081 gttccgttcc gctatcctgc cggcgattat cacgctgatc ttttaccagg gagttaagcc
     1141 actcatgctg ttaaatgcgc tggcgtcgct cggacataaa gggattaaaa ccctgagaac
     1201 gttcgggcgg gccgggttaa tgttattcaa tgcgctggtc ggcaaaccgg aatttcgcaa
     1261 acatgcgccg ctgctggtgc gccagctcta taatgtcggc gtcctgtcga tgctgattat
     1321 tgtggtttct ggcgtgttca tcggaatggt gttggggctg caaggttatc tggttctgac
     1381 cacttatagt gcggaaacca gtctgggtat gctggtggcg ttatcgctac tgcgtgaact
     1441 ggggccggtg gttgccgcgt tgttgtttgc cgggcgtgct ggttcggcgc taaccgcaga
     1501 aatcggcctg atgcgcgcta cagagcaact ctccagtatg gagatgatgg cggtggatcc
     1561 gctgcgtcgg gttatttctc cccgtttctg ggctggggtt atttcattac cactgttgac
     1621 ggttattttc gtcgccgtgg ggatctgggg cggatctctg gtcggcgtca gttggaaagg
     1681 cattgatagc gggttcttct ggtcggcaat gcaaaatgcc gtcgactggc gtatggatct
     1741 ggtcaactgt ctgattaaga gcgtggtgtt cgccatcacg gtgacgtgga tttcgttgtt
     1801 taacggctac gacgccatcc cgacgtctgc cgggattagc cgggcaacca ctcgcaccgt
     1861 tgtccactcg tctctggctg ttctggggct ggattttgtg ctgaccgcat tgatgtttgg
     1921 gaattgagtt catgcaaacg aaaaaaaatg aaatttgggt gggtatcttt ttattagcag
     1981 cactgctggc ggcgctgttt gtttgcctga aggcggcgaa cgtgacgtcc atacgtactg
     2041 aaccgaccta cacgctttat gcgacgttcg ataacattgg cggcctgaaa gcgcgctctc
     2101 cggtcagtat tggtggcgtt gttgtgggtc gggtggcgga tattacgctg gacccgaaaa
     2161 cctatctgcc gcgcgtaacg ctggaaattg aacaacgtta taaccacatt ccagatacca
     2221 gttcgctgag cattcgtact tccggcctgc tgggggaaca atatctggca ttaaacgtcg
     2281 gttttgaaga cccggaactg gggactgcta tcctgaagga tggcgataca attcaggaca
     2341 ctaagtctgc gtaatttcag gagaaccgac gcatgtttaa acgtttaatg atggtcgctt
     2401 tgctggtgat tgcacctctg agtgcggcaa ccgcggcaga ccagaccaat ccgtataagc
     2461 tgatggacga ggcggcgcag aaaacgttcg atcgcctgaa gaatgagcaa ccgcaaattc
     2521 gggccaaccc ggattatctg cgtaccattg ttgatcagga actgctgcca tacgtacagg
     2581 tgaaatacgc cggtgcgctg gtgctgggcc agtattacaa gagtgcgacc cctgctcaac
     2641 gtgaagccta ctttgccgct ttccgtgagt acctgaagca ggcttacggt caggcgctgg
     2701 cgatgtatca cggtcaaacc tatcagattg cgccagaaca gccgctgggc gataaaacca
     2761 ttgtgcctat tcgcgttacc attattgacc cgaatggccg tccgccggtg cgtctggact
     2821 tccagtggcg taaaaactcc cagacgggca attggcaggc ttacgacatg attgctgaag
     2881 gcgtcagtat gatcaccacc aaacaaaacg agtggggaac gctgctgcgt accaaaggta
     2941 tcgacggcct gactgcgcaa ctgaaatcga tttctcaaca gaaaatcact ctggaagaga
     3001 aaaaataatg agcgagtcac tgagctggat gcagacgggt gacacgctgg cgttatccgg
     3061 agagctggat caggacgttt tgctaccgct ttgggaaatg cgtgaggaag cggtgaaggg
     3121 gattacctgc atcgatctta gccgtgtctc ccgcgtggat acggggggac tggcactgct
     3181 gctccatctt atcgatctgg cgaaaaagca gggcaacaat gtgacgcttc agggggtaaa
     3241 cgacaaagtg tataccctgg cgaaattgta taatttgcct gctgatgttc tgcctcgtta
     3301 aGGTACCAAA TTCCAGAAAA GAGACGCTGA AAAGCGTCTT TTTTCGTTTT GGTCCTGGTC
     3361 TGAGGAATGG TTGTAAGCTT GGCTGTTTTG GCGGATGAGA GAAGATTTTC AGCCTGATAC
     3421 AGATTAAATC AGAACGCAGA AGCGGTCTGA TAAAACAGAA TTTGCCTGGC GGCAGTAGCG
     3481 CGGTGGTCCC ACCTGACCCC ATGCCGAACT CAGAAGTGAA ACGCCGTAGC GCCGATGGTA
     3541 GTGTGGGGTC TCCCCATGCG AGAGTAGGGA ACTGCCAGGC ATCAAATAAA ACGAAAGGCT
     3601 CAGTCGAAAG ACTGGGCCTT TCGTTTTATC TGTTGTTTGT CGGTGAACGC TCTCCTGAGT
     3661 AGGACAAATC CGCCGGGAGC GGATTTGAAC GTTGCGAAGC AACGGCCCGG AGGGTGGCGG
     3721 GCAGGACGCC CGCCATAAAC TGCCAGGCAT CAAATTAAGC AGAAGGCCAT CCTGACGGAT
     3781 GGCCTTTTTG CGTTTCTACA AACTCTTTTG TTTATTTTTC TAAATACATT CAAATATGTA
     3841 TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA GGAAGAGTAT
     3901 GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT GCCTTCCTGT
     3961 TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT TGGGTGCACG
     4021 AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT TTCGCCCCGA
     4081 AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG TATTATCCCG
     4141 TGTTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA ATGACTTGGT
     4201 TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA GAGAATTATG
     4261 CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA CAACGATCGG
     4321 AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA CTCGCCTTGA
     4381 TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA CCACGATGCC
     4441 TGcAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA CTCTAGCTTC
     4501 CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC TTCTGCGCTC
     4561 GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC GTGGGTCTCG
     4621 CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG TTATCTACAC
     4681 GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA TAGGTGCCTC
     4741 ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT AGATTGATTT
     4801 ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG CGGGTGTGGT GGTTACGCGC AGCGTGACCG
     4861 CTACACTTGC CAGCGCCCTA GCGCCCGCTC CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA
     4921 CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA ATCGGGGGCT CCCTTTAGGG TTCCGATTTA
     4981 GTGCTTTACG GCACCTCGAC CCCAAAAAAC TTGATTTGGG TGATGGTTCA CGTAGTGGGC
     5041 CATCGCCCTG ATAGACGGTT TTTCGCCCTT TGACGTTGGA GTCCACGTTC TTTAATAGTG
     5101 GACTCTTGTT CCAAACTGGA ACAACACTCA ACCCTATCTC GGGCTATTCT TTTGATTTAT
     5161 AAGGGATTTT GCCGATTTCG GCCTATTGGT TAAAAAATGA GCTGATTTAA CAAAAATTTA
     5221 ACGCGAATTT TAACAAAATA TTAACGTTTA CAATTTAAAA GGATCTAGGT GAAGATCCTT
     5281 TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC
     5341 CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC
     5401 TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA
     5461 ACTCTTTTTC CGAAGGTAAC TGGCTCAGCA GAGCGCAGAT ACCAAATACT GTCCTTCTAG
     5521 TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC
     5581 TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG
     5641 ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA
     5701 CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCTAT
     5761 GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG
     5821 TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC
     5881 CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC
     5941 GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC
     6001 CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC CGTATTACCG
     6061 CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC CGAGCGCAGC GAGTCAGTGA
     6121 GCGAGGAAGC GGAAGAGCGC CTGATGCGGT ATTTTCTCCT TACGCATCTG TGCGGTATTT
     6181 CACACCGCAT AGGGTCATGG CTGCGCCCCG ACACCCGCCA ACACCCGCTG ACGCGCCCTG
     6241 ACGGGCTTGT CTGCTCCCGG CATCCGCTTA CAGACAAGCT GTGACCGTCT CCGGGAGCTG
     6301 CATGTGTCAG AGGTTTTCAC CGTCATCACC GAAACGCGCG AGGCAGaAGg agATGGCGCC
     6361 CAACAGTCCC CCGGCCACGG GGCCTGCCAC CATACCCACG CCGAAACAAG CGCTCATGAG
     6421 CCCGAAGTGG CGAGCCCGAT CTTCCCCATC GGTGATGTCG GCGATATAGG CGCCAGCAAC
     6481 CGCACCTGTG GCGCCGGTGA TGCCGGCCAC GATGCGTCCG GCGTAGAGGA TCTGCTCATG
     6541 TTTGACAGCT TATCATCGAT GCATAATGTG CCTGTCAAAT GGACGAAGCA GGGATTCTGC
     6601 AAACCCTATG CTACTCCGTC AAGCCGTCAA TTGTCTGATT CGTTACCAAT TATGACAACT
     6661 TGACGGCTAC ATCATTCACT TTTTCTTCAC AACCGGCACG GAACTCGCTC GGGCTGGCCC
     6721 CGGTGCATTT TTTAAATACC CGCGAGAAAT AGAGTTGATC GTCAAAACCA ACATTGCGAC
     6781 CGACGGTGGC GATAGGCATC CGGGTGGTGC TCAAAAGCAG CTTCGCCTGG CTGATACGTT
     6841 GGTCCTCGCG CCAGCTTAAG ACGCTAATCC CTAACTGCTG GCGGAAAAGA TGTGACAGAC
     6901 GCGACGGCGA CAAGCAAACA TGCTGTGCGA CGCTGGCGAT ATCAAAATTG CTGTCTGCCA
     6961 GGTGATCGCT GATGTACTGA CAAGCCTCGC GTACCCGATT ATCCATCGGT GGATGGAGCG
     7021 ACTCGTTAAT CGCTTCCATG CGCCGCAGTA ACAATTGCTC AAGCAGATTT ATCGCCAGCA
     7081 GCTCCGAATA GCGCCCTTCC CCTTGCCCGG CGTTAATGAT TTGCCCAAAC AGGTCGCTGA
     7141 AATGCGGCTG GTGCGCTTCA TCCGGGCGAA AGAACCCCGT ATTGGCAAAT ATTGACGGCC
     7201 AGTTAAGCCA TTCATGCCAG TAGGCGCGCG GACGAAAGTA AACCCACTGG TGATACCATT
     7261 CGCGAGCCTC CGGATGACGA CCGTAGTGAT GAATCTCTCC TGGCGGGAAC AGCAAAATAT
     7321 CACCCGGTCG GCAAACAAAT TCTCGTCCCT GATTTTTCAC CACCCCCTGA CCGCGAATGG
     7381 TGAGATTGAG AATATAACCT TTCATTCCCA GCGGTCGGTC GATAAAAAAA TCGAGATAAC
     7441 CGTTGGCCTC AATCGGCGTT AAACCCGCCA CCAGATGGGC ATTAAACGAG TATCCCGGCA
     7501 GCAGGGGATC ATTTTGCGCT TCAGCCATAC TTTTCATACT CCCGC
//