LOCUS       sequence-320182-        7773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaD TM swap with LetB 
            TM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7773)
  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 7773)
  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: pBEL2138
FEATURES             Location/Qualifiers
     source          1..7773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..337
                     /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     260..279
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1191..1208
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1221..1241
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3684..3701)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3854..3940
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4032..4059
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4078..4169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4170..4238,4239..5030)
                     /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(4388..4407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5072..5527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5159..5178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5369..5390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5638..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6379..6396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6562..6584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6722..6741
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6921..7773,1..26))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag agaagaaacc
       61 aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct cttctcgcta
      121 accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac caaagccatg
      181 acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat tgattatttg
      241 cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc ggatcctacc
      301 tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg ggctagccga
      361 agggtgaatt atggagcagt ctgtggcgaa tttagtcgat atgcgcgatg tcagttttac
      421 gcgtggcaat cgctgcatct tcgataatat ttccctgacc gtgccgcgag ggaagatcac
      481 ggcgatcatg gggccatcgg gcatcggtaa aacgacgcta ctccgtctga ttggcgggca
      541 aatcgcacca gatcatggtg agatcctttt cgatggtgag aatattccgg cgatgtctcg
      601 ttcgcgcctg tatacagtgc gcaaacggat gagcatgtta tttcagtccg gggcgttgtt
      661 cactgatatg aacgtatttg acaacgtcgc ctatccactg cgcgaacata cccaacttcc
      721 cgcgccattg ttgcatagta cggtgatgat gaagctggag gccgtggggc tgcgtggagc
      781 ggctaaacta atgccttctg aactttccgg tgggatggcg cggcgtgcag cgctggcgcg
      841 tgcgattgcg ctggagccgg atctcatcat gtttgatgaa ccttttgttg ggcaagatcc
      901 catcaccatg ggcgtactgg tgaagctgat ttctgagctg aacagcgcgc tgggcgtgac
      961 ttgtgtggtg gtttctcacg atgtgccgga agtgttaagt attgcggatc acgcctggat
     1021 cctggcggac aaaaaaattg tcgctcatgg cagtgcccag gcgttgcagg cgaatcctga
     1081 tccgcgcgta cgtcagtttc tggacgggat agctgacggg cctgttccgt tccgctatcc
     1141 tgccggcgat tatcacgctg atcttttacc agggagttaa gccactcatg catcaccacc
     1201 atcaccatca gcatcaacac gaaaatctgt attttcaggg gatgctgtta aatgcgctgg
     1261 cgtcgctcgg acataaaggg attaaaaccc tgagaacgtt cgggcgggcc gggttaatgt
     1321 tattcaatgc gctggtcggc aaaccggaat ttcgcaaaca tgcgccgctg ctggtgcgcc
     1381 agctctataa tgtcggcgtc ctgtcgatgc tgattattgt ggtttctggc gtgttcatcg
     1441 gaatggtgtt ggggctgcaa ggttatctgg ttctgaccac ttatagtgcg gaaaccagtc
     1501 tgggtatgct ggtggcgtta tcgctactgc gtgaactggg gccggtggtt gccgcgttgt
     1561 tgtttgccgg gcgtgctggt tcggcgctaa ccgcagaaat cggcctgatg cgcgctacag
     1621 agcaactctc cagtatggag atgatggcgg tggatccgct gcgtcgggtt atttctcccc
     1681 gtttctgggc tggggttatt tcattaccac tgttgacggt tattttcgtc gccgtgggga
     1741 tctggggcgg atctctggtc ggcgtcagtt ggaaaggcat tgatagcggg ttcttctggt
     1801 cggcaatgca aaatgccgtc gactggcgta tggatctggt caactgtctg attaagagcg
     1861 tggtgttcgc catcacggtg acgtggattt cgttgtttaa cggctacgac gccatcccga
     1921 cgtctgccgg gattagccgg gcaaccactc gcaccgttgt ccactcgtct ctggctgttc
     1981 tggggctgga ttttgtgctg accgcattga tgtttgggaa ttgagttcat gagtcaggaa
     2041 acgcccgctt cgacgactga agcgcagatt aaaaataaac gccgtatctc acctttctgg
     2101 ctgctgccgt tcatcgcgct aatgattgcc agttggctga tttgggacag ttatcaggcg
     2161 aacgtgacgt ccatacgtac tgaaccgacc tacacgcttt atgcgacgtt cgataacatt
     2221 ggcggcctga aagcgcgctc tccggtcagt attggtggcg ttgttgtggg tcgggtggcg
     2281 gatattacgc tggacccgaa aacctatctg ccgcgcgtaa cgctggaaat tgaacaacgt
     2341 tataaccaca ttccagatac cagttcgctg agcattcgta cttccggcct gctgggggaa
     2401 caatatctgg cattaaacgt cggttttgaa gacccggaac tggggactgc tatcctgaag
     2461 gatggcgata caattcagga cactaagtct gcgatggtgc tggaagatct cattggtcag
     2521 ttcctttacg gtagtaaagg cgatgacaat aagaatagtg gcgatgcgcc agctgctgcg
     2581 ccaggtaata atgaaaccac tgaacctgtg ggtacaacga aataatttca ggagaaccga
     2641 cgcatgttta aacgtttaat gatggtcgct ttgctggtga ttgcacctct gagtgcggca
     2701 accgcggcag accagaccaa tccgtataag ctgatggacg aggcggcgca gaaaacgttc
     2761 gatcgcctga agaatgagca accgcaaatt cgggccaacc cggattatct gcgtaccatt
     2821 gttgatcagg aactgctgcc atacgtacag gtgaaatacg ccggtgcgct ggtgctgggc
     2881 cagtattaca agagtgcgac ccctgctcaa cgtgaagcct actttgccgc tttccgtgag
     2941 tacctgaagc aggcttacgg tcaggcgctg gcgatgtatc acggtcaaac ctatcagatt
     3001 gcgccagaac agccgctggg cgataaaacc attgtgccta ttcgcgttac cattattgac
     3061 ccgaatggcc gtccgccggt gcgtctggac ttccagtggc gtaaaaactc ccagacgggc
     3121 aattggcagg cttacgacat gattgctgaa ggcgtcagta tgatcaccac caaacaaaac
     3181 gagtggggaa cgctgctgcg taccaaaggt atcgacggcc tgactgcgca actgaaatcg
     3241 atttctcaac agaaaatcac tctggaagag aaaaaataat gagcgagtca ctgagctgga
     3301 tgcagacggg tgacacgctg gcgttatccg gagagctgga tcaggacgtt ttgctaccgc
     3361 tttgggaaat gcgtgaggaa gcggtgaagg ggattacctg catcgatctt agccgtgtct
     3421 cccgcgtgga tacgggggga ctggcactgc tgctccatct tatcgatctg gcgaaaaagc
     3481 agggcaacaa tgtgacgctt cagggggtaa acgacaaagt gtataccctg gcgaaattgt
     3541 ataatttgcc tgctgatgtt ctgcctcgtt aaggtaccaa attccagaaa agagacgctg
     3601 aaaagcgtct tttttcgttt tggtcctggt ctgaggaatg gttgtaagct tggctgtttt
     3661 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     3721 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     3781 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     3841 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3901 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     3961 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     4021 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     4081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4141 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4261 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4321 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4381 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4561 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4681 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     4741 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4861 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5041 aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg
     5101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5281 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5401 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5521 acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggctcagc
     5761 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6301 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6361 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     6421 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtcatg gctgcgcccc
     6481 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6541 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6601 cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6661 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6721 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6781 cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcatcga tgcataatgt
     6841 gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca
     6901 attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca
     6961 caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa
     7021 tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg
     7081 ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc
     7141 cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg
     7201 acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg
     7261 cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt
     7321 aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg
     7381 gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga
     7441 aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc
     7501 ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga
     7561 tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc
     7621 tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc
     7681 agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc
     7741 accagatggg cattaaacga gtatcccggc agc
//
