LOCUS       sequence-320180-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for MlaE E98A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7728)
  AUTHORS   Coudray N, Isom GL, MacRae MR, Saiduddin MN, Bhabha G, Ekiert DC
  TITLE     Structure of bacterial phospholipid transporter MlaFEDB with 
            substrate bound.
  JOURNAL   Elife. 2020 Nov 25;9. pii: 62518. doi: 10.7554/eLife.62518.
  PUBMED    33236984
REFERENCE   2  (bases 1 to 7728)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2020-11-25"; volume: "9. pii: 62518. doi"; pages: " 
            10.7554/eLife.62518"
COMMENT     Sequence Label: pBEL2094
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..343
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     266..285
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1984..2001
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2014..2034
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3645..3662)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3815..3901
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3993..4020
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4039..4130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4131..4199,4200..4991)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4349..4368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5033..5488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5120..5139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5330..5351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5599..6186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6087..6106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6340..6357
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6523..6545
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6683..6702
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6882..7728,1..32))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa
       61 gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc
      121 tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa
      181 gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat
      241 tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat
      301 cctacctgac gctttttatc gcaactctct actgtttctc catacccgtt tttttgggct
      361 agccgaaggg tgaattatgg agcagtctgt ggcgaattta gtcgatatgc gcgatgtcag
      421 ttttacgcgt ggcaatcgct gcatcttcga taatatttcc ctgaccgtgc cgcgagggaa
      481 gatcacggcg atcatggggc catcgggcat cggtaaaacg acgctactcc gtctgattgg
      541 cgggcaaatc gcaccagatc atggtgagat ccttttcgat ggtgagaata ttccggcgat
      601 gtctcgttcg cgcctgtata cagtgcgcaa acggatgagc atgttatttc agtccggggc
      661 gttgttcact gatatgaacg tatttgacaa cgtcgcctat ccactgcgcg aacataccca
      721 acttcccgcg ccattgttgc atagtacggt gatgatgaag ctggaggccg tggggctgcg
      781 tggagcggct aaactaatgc cttctgaact ttccggtggg atggcgcggc gtgcagcgct
      841 ggcgcgtgcg attgcgctgg agccggatct catcatgttt gatgaacctt ttgttgggca
      901 agatcccatc accatgggcg tactggtgaa gctgatttct gagctgaaca gcgcgctggg
      961 cgtgacttgt gtggtggttt ctcacgatgt gccggaagtg ttaagtattg cggatcacgc
     1021 ctggatcctg gcggacaaaa aaattgtcgc tcatggcagt gcccaggcgt tgcaggcgaa
     1081 tcctgatccg cgcgtacgtc agtttctgga cgggatagct gacgggcctg ttccgttccg
     1141 ctatcctgcc ggcgattatc acgctgatct tttaccaggg agttaagcca ctcatgctgt
     1201 taaatgcgct ggcgtcgctc ggacataaag ggattaaaac cctgagaacg ttcgggcggg
     1261 ccgggttaat gttattcaat gcgctggtcg gcaaaccgga atttcgcaaa catgcgccgc
     1321 tgctggtgcg ccagctctat aatgtcggcg tcctgtcgat gctgattatt gtggtttctg
     1381 gcgtgttcat cggaatggtg ttggggctgc aaggttatct ggttctgacc acttatagtg
     1441 cggaaaccag tctgggtatg ctggtggcgt tatcgctact gcgtgcgctg gggccggtgg
     1501 ttgccgcgtt gttgtttgcc gggcgtgctg gttcggcgct aaccgcagaa atcggcctga
     1561 tgcgcgctac agagcaactc tccagtatgg agatgatggc ggtggatccg ctgcgtcggg
     1621 ttatttctcc ccgtttctgg gctggggtta tttcattacc actgttgacg gttattttcg
     1681 tcgccgtggg gatctggggc ggatctctgg tcggcgtcag ttggaaaggc attgatagcg
     1741 ggttcttctg gtcggcaatg caaaatgccg tcgactggcg tatggatctg gtcaactgtc
     1801 tgattaagag cgtggtgttc gccatcacgg tgacgtggat ttcgttgttt aacggctacg
     1861 acgccatccc gacgtctgcc gggattagcc gggcaaccac tcgcaccgtt gtccactcgt
     1921 ctctggctgt tctggggctg gattttgtgc tgaccgcatt gatgtttggg aattgagttc
     1981 atgcatcacc accatcacca tcagcatcaa cacgaaaatc tgtattttca ggggatgcaa
     2041 acgaaaaaaa atgaaatttg ggtgggtatc tttttattag cagcactgct ggcggcgctg
     2101 tttgtttgcc tgaaggcggc gaacgtgacg tccatacgta ctgaaccgac ctacacgctt
     2161 tatgcgacgt tcgataacat tggcggcctg aaagcgcgct ctccggtcag tattggtggc
     2221 gttgttgtgg gtcgggtggc ggatattacg ctggacccga aaacctatct gccgcgcgta
     2281 acgctggaaa ttgaacaacg ttataaccac attccagata ccagttcgct gagcattcgt
     2341 acttccggcc tgctggggga acaatatctg gcattaaacg tcggttttga agacccggaa
     2401 ctggggactg ctatcctgaa ggatggcgat acaattcagg acactaagtc tgcgatggtg
     2461 ctggaagatc tcattggtca gttcctttac ggtagtaaag gcgatgacaa taagaatagt
     2521 ggcgatgcgc cagctgctgc gccaggtaat aatgaaacca ctgaacctgt gggtacaacg
     2581 aaataatttc aggagaaccg acgcatgttt aaacgtttaa tgatggtcgc tttgctggtg
     2641 attgcacctc tgagtgcggc aaccgcggca gaccagacca atccgtataa gctgatggac
     2701 gaggcggcgc agaaaacgtt cgatcgcctg aagaatgagc aaccgcaaat tcgggccaac
     2761 ccggattatc tgcgtaccat tgttgatcag gaactgctgc catacgtaca ggtgaaatac
     2821 gccggtgcgc tggtgctggg ccagtattac aagagtgcga cccctgctca acgtgaagcc
     2881 tactttgccg ctttccgtga gtacctgaag caggcttacg gtcaggcgct ggcgatgtat
     2941 cacggtcaaa cctatcagat tgcgccagaa cagccgctgg gcgataaaac cattgtgcct
     3001 attcgcgtta ccattattga cccgaatggc cgtccgccgg tgcgtctgga cttccagtgg
     3061 cgtaaaaact cccagacggg caattggcag gcttacgaca tgattgctga aggcgtcagt
     3121 atgatcacca ccaaacaaaa cgagtgggga acgctgctgc gtaccaaagg tatcgacggc
     3181 ctgactgcgc aactgaaatc gatttctcaa cagaaaatca ctctggaaga gaaaaaataa
     3241 tgagcgagtc actgagctgg atgcagacgg gtgacacgct ggcgttatcc ggagagctgg
     3301 atcaggacgt tttgctaccg ctttgggaaa tgcgtgagga agcggtgaag gggattacct
     3361 gcatcgatct tagccgtgtc tcccgcgtgg atacgggggg actggcactg ctgctccatc
     3421 ttatcgatct ggcgaaaaag cagggcaaca atgtgacgct tcagggggta aacgacaaag
     3481 tgtataccct ggcgaaattg tataatttgc ctgctgatgt tctgcctcgt taaggtacca
     3541 aattccagaa aagagacgct gaaaagcgtc ttttttcgtt ttggtcctgg tctgaggaat
     3601 ggttgtaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     3661 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     3721 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     3781 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     3841 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     3901 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     3961 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     4021 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4081 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4141 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4201 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4261 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4321 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     4381 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4441 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4501 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4561 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4621 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     4681 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4741 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4801 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4861 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4921 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4981 agcattggta actgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     5041 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5101 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5161 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5221 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     5281 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5341 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     5401 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5461 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     5521 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5581 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5641 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5701 tccgaaggta actggctcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6301 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6361 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6421 atagggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6481 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6541 agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc
     6601 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6661 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6721 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag
     6781 cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta
     6841 tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct
     6901 acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat
     6961 tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg
     7021 gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg
     7081 cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc
     7141 gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg
     7201 ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta
     7261 atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa
     7321 tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc
     7381 tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc
     7441 cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc
     7501 tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt
     7561 cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg
     7621 agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc
     7681 tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatccc
//
