LOCUS       sequence-326368-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for MlaD (F13A).
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: pBEL2294
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(38..625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(118..137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(736..1191)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(873..894)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1085..1104
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(1233..2024,2025..2093))
                     /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     1856..1875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2094..2185)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2204..2231)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2323..2409)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2562..2579
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4977..4997)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5010..5027)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(5881..6165)
                     /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     complement(5939..5958)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6192..7070
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(7250..7269)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7407..7429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7595..7612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
       61 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
      481 cagtatttgg tatctgcgct ctgctgagcc agttaccttc ggaaaaagag ttggtagctc
      541 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      601 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      661 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      721 cacctagatc cttttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
      781 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      841 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
      901 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
      961 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     1021 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1081 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     1141 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg taaatcaatc
     1201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     1561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     1861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2161 gaatgtattt agaaaaataa acaaaagagt ttgtagaaac gcaaaaaggc catccgtcag
     2221 gatggccttc tgcttaattt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     2281 tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     2341 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     2401 ttttatttga tgcctggcag ttccctactc tcgcatgggg agaccccaca ctaccatcgg
     2461 cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc gcgctactgc
     2521 cgccaggcaa attctgtttt atcagaccgc ttctgcgttc tgatttaatc tgtatcaggc
     2581 tgaaaatctt ctctcatccg ccaaaacagc caagcttaca accattcctc agaccaggac
     2641 caaaacgaaa aaagacgctt ttcagcgtct cttttctgga atttggtacc ttaacgaggc
     2701 agaacatcag caggcaaatt atacaatttc gccagggtat acactttgtc gtttaccccc
     2761 tgaagcgtca cattgttgcc ctgctttttc gccagatcga taagatggag cagcagtgcc
     2821 agtccccccg tatccacgcg ggagacacgg ctaagatcga tgcaggtaat ccccttcacc
     2881 gcttcctcac gcatttccca aagcggtagc aaaacgtcct gatccagctc tccggataac
     2941 gccagcgtgt cacccgtctg catccagctc agtgactcgc tcattatttt ttctcttcca
     3001 gagtgatttt ctgttgagaa atcgatttca gttgcgcagt caggccgtcg atacctttgg
     3061 tacgcagcag cgttccccac tcgttttgtt tggtggtgat catactgacg ccttcagcaa
     3121 tcatgtcgta agcctgccaa ttgcccgtct gggagttttt acgccactgg aagtccagac
     3181 gcaccggcgg acggccattc gggtcaataa tggtaacgcg aataggcaca atggttttat
     3241 cgcccagcgg ctgttctggc gcaatctgat aggtttgacc gtgatacatc gccagcgcct
     3301 gaccgtaagc ctgcttcagg tactcacgga aagcggcaaa gtaggcttca cgttgagcag
     3361 gggtcgcact cttgtaatac tggcccagca ccagcgcacc ggcgtatttc acctgtacgt
     3421 atggcagcag ttcctgatca acaatggtac gcagataatc cgggttggcc cgaatttgcg
     3481 gttgctcatt cttcaggcga tcgaacgttt tctgcgccgc ctcgtccatc agcttatacg
     3541 gattggtctg gtctgccgcg gttgccgcac tcagaggtgc aatcaccagc aaagcgacca
     3601 tcattaaacg tttaaacatg cgtcggttct cctgaaatta tttcgttgta cccacaggtt
     3661 cagtggtttc attattacct ggcgcagcag ctggcgcatc gccactattc ttattgtcat
     3721 cgcctttact accgtaaagg aactgaccaa tgagatcttc cagcaccatc gcagacttag
     3781 tgtcctgaat tgtatcgcca tccttcagga tagcagtccc cagttccggg tcttcaaaac
     3841 cgacgtttaa tgccagatat tgttccccca gcaggccgga agtacgaatg ctcagcgaac
     3901 tggtatctgg aatgtggtta taacgttgtt caatttccag cgttacgcgc ggcagatagg
     3961 ttttcgggtc cagcgtaata tccgccaccc gacccacaac aacgccacca atactgaccg
     4021 gagagcgcgc tttcaggccg ccaatgttat cgaacgtcgc ataaagcgtg taggtcggtt
     4081 cagtacgtat ggacgtcacg ttcgccgcct tcaggcaaac aaacagcgcc gccagcagtg
     4141 ctgctaataa cgcgataccc acccaaattt catttttttt cgtttgcatg aactcaattc
     4201 ccaaacatca atgcggtcag cacaaaatcc agccccagaa cagccagaga cgagtggaca
     4261 acggtgcgag tggttgcccg gctaatcccg gcagacgtcg ggatggcgtc gtagccgtta
     4321 aacaacgaaa tccacgtcac cgtgatggcg aacaccacgc tcttaatcag acagttgacc
     4381 agatccatac gccagtcgac ggcattttgc attgccgacc agaagaaccc gctatcaatg
     4441 cctttccaac tgacgccgac cagagatccg ccccagatcc ccacggcgac gaaaataacc
     4501 gtcaacagtg gtaatgaaat aaccccagcc cagaaacggg gagaaataac ccgacgcagc
     4561 ggatccaccg ccatcatctc catactggag agttgctctg tagcgcgcat caggccgatt
     4621 tctgcggtta gcgccgaacc agcacgcccg gcaaacaaca acgcggcaac caccggcccc
     4681 agttcacgca gtagcgataa cgccaccagc atacccagac tggtttccgc actataagtg
     4741 gtcagaacca gataaccttg cagccccaac accattccga tgaacacgcc agaaaccaca
     4801 ataatcagca tcgacaggac gccgacatta tagagctggc gcaccagcag cggcgcatgt
     4861 ttgcgaaatt ccggtttgcc gaccagcgca ttgaataaca ttaacccggc ccgcccgaac
     4921 gttctcaggg ttttaatccc tttatgtccg agcgacgcca gcgcatttaa cagcatcccc
     4981 tgaaaataca gattttcgtg ttgatgctga tggtgatggt ggtgatgcat gagtggctta
     5041 actccctggt aaaagatcag cgtgataatc gccggcagga tagcggaacg gaacaggccc
     5101 gtcagctatc ccgtccagaa actgacgtac gcgcggatca ggattcgcct gcaacgcctg
     5161 ggcactgcca tgagcgacaa tttttttgtc cgccaggatc caggcgtgat ccgcaatact
     5221 taacacttcc ggcacatcgt gagaaaccac cacacaagtc acgcccagcg cgctgttcag
     5281 ctcagaaatc agcttcacca gtacgcccat ggtgatggga tcttgcccaa caaaaggttc
     5341 atcaaacatg atgagatccg gctccagcgc aatcgcacgc gccagcgctg cacgccgcgc
     5401 catcccaccg gaaagttcag aaggcattag tttagccgct ccacgcagcc ccacggcctc
     5461 cagcttcatc atcaccgtac tatgcaacaa tggcgcggga agttgggtat gttcgcgcag
     5521 tggataggcg acgttgtcaa atacgttcat atcagtgaac aacgccccgg actgaaataa
     5581 catgctcatc cgtttgcgca ctgtatacag gcgcgaacga gacatcgccg gaatattctc
     5641 accatcgaaa aggatctcac catgatctgg tgcgatttgc ccgccaatca gacggagtag
     5701 cgtcgtttta ccgatgcccg atggccccat gatcgccgtg atcttccctc gcggcacggt
     5761 cagggaaata ttatcgaaga tgcagcgatt gccacgcgta aaactgacat cgcgcatatc
     5821 gactaaattc gccacagact gctccataat tcacccttcg gctagcccaa aaaaacgggt
     5881 atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct aatcttatgg
     5941 ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg tggacttttc
     6001 tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc ccgctttgtt
     6061 acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc cagtaaaaga
     6121 cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg aatggcggga
     6181 gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac tcgtttaatg
     6241 cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat ttttttatcg
     6301 accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt cagggggtgg
     6361 tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg ttcccgccag
     6421 gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac cagtgggttt
     6481 actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata tttgccaata
     6541 cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg tttgggcaaa
     6601 tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata aatctgcttg
     6661 agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca ccgatggata
     6721 atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc aattttgata
     6781 tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat cttttccgcc
     6841 agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc caggcgaagc
     6901 tgcttttgag caccacccgg atgcctatcg ccaccgtcgg tcgcaatgtt ggttttgacg
     6961 atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg ggccagcccg agcgagttcc
     7021 gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa gttgtcataa ttggtaacga
     7081 atcagacaat tgacggcttg acggagtagc atagggtttg cagaatccct gcttcgtcca
     7141 tttgacaggc acattatgca tcgatgataa gctgtcaaac atgagcagat cctctacgcc
     7201 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     7261 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     7321 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttctgcctc
     7381 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7441 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     7501 ggcgggtgtc ggggcgcagc catgacccta tgcggtgtga aataccgcac agatgcgtaa
     7561 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7621 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7681 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaa
//
