LOCUS       sequence-320141-        7716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaD TM swap with LptC 
            TM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7716)
  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 7716)
  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: pBEL2139
FEATURES             Location/Qualifiers
     source          1..7716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..294
                     /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     217..236
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1148..1165
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1178..1198
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3584..3601)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3754..3840
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3932..3959
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3978..4069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4070..4138,4139..4930)
                     /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(4288..4307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4972..5427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5059..5078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5269..5290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5538..6125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6026..6045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6279..6296
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6462..6484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6622..6641
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6821..7699)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
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 actcatgcat caccaccatc accatcagca tcaacacgaa aatctgtatt ttcaggggat
     1201 gctgttaaat gcgctggcgt cgctcggaca taaagggatt aaaaccctga gaacgttcgg
     1261 gcgggccggg ttaatgttat tcaatgcgct ggtcggcaaa ccggaatttc gcaaacatgc
     1321 gccgctgctg gtgcgccagc tctataatgt cggcgtcctg tcgatgctga ttattgtggt
     1381 ttctggcgtg ttcatcggaa tggtgttggg gctgcaaggt tatctggttc tgaccactta
     1441 tagtgcggaa accagtctgg gtatgctggt ggcgttatcg ctactgcgtg aactggggcc
     1501 ggtggttgcc gcgttgttgt ttgccgggcg tgctggttcg gcgctaaccg cagaaatcgg
     1561 cctgatgcgc gctacagagc aactctccag tatggagatg atggcggtgg atccgctgcg
     1621 tcgggttatt tctccccgtt tctgggctgg ggttatttca ttaccactgt tgacggttat
     1681 tttcgtcgcc gtggggatct ggggcggatc tctggtcggc gtcagttgga aaggcattga
     1741 tagcgggttc ttctggtcgg caatgcaaaa tgccgtcgac tggcgtatgg atctggtcaa
     1801 ctgtctgatt aagagcgtgg tgttcgccat cacggtgacg tggatttcgt tgtttaacgg
     1861 ctacgacgcc atcccgacgt ctgccgggat tagccgggca accactcgca ccgttgtcca
     1921 ctcgtctctg gctgttctgg ggctggattt tgtgctgacc gcattgatgt ttgggaattg
     1981 agttcatgag taaagccaga cgttgggtta tcattgtgct atcactggcg gttctggtga
     2041 tgatcggcat taatatggcg aacgtgacgt ccatacgtac tgaaccgacc tacacgcttt
     2101 atgcgacgtt cgataacatt ggcggcctga aagcgcgctc tccggtcagt attggtggcg
     2161 ttgttgtggg tcgggtggcg gatattacgc tggacccgaa aacctatctg ccgcgcgtaa
     2221 cgctggaaat tgaacaacgt tataaccaca ttccagatac cagttcgctg agcattcgta
     2281 cttccggcct gctgggggaa caatatctgg cattaaacgt cggttttgaa gacccggaac
     2341 tggggactgc tatcctgaag gatggcgata caattcagga cactaagtct gcgatggtgc
     2401 tggaagatct cattggtcag ttcctttacg gtagtaaagg cgatgacaat aagaatagtg
     2461 gcgatgcgcc agctgctgcg ccaggtaata atgaaaccac tgaacctgtg ggtacaacga
     2521 aataatttca ggagaaccga cgcatgttta aacgtttaat gatggtcgct ttgctggtga
     2581 ttgcacctct gagtgcggca accgcggcag accagaccaa tccgtataag ctgatggacg
     2641 aggcggcgca gaaaacgttc gatcgcctga agaatgagca accgcaaatt cgggccaacc
     2701 cggattatct gcgtaccatt gttgatcagg aactgctgcc atacgtacag gtgaaatacg
     2761 ccggtgcgct ggtgctgggc cagtattaca agagtgcgac ccctgctcaa cgtgaagcct
     2821 actttgccgc tttccgtgag tacctgaagc aggcttacgg tcaggcgctg gcgatgtatc
     2881 acggtcaaac ctatcagatt gcgccagaac agccgctggg cgataaaacc attgtgccta
     2941 ttcgcgttac cattattgac ccgaatggcc gtccgccggt gcgtctggac ttccagtggc
     3001 gtaaaaactc ccagacgggc aattggcagg cttacgacat gattgctgaa ggcgtcagta
     3061 tgatcaccac caaacaaaac gagtggggaa cgctgctgcg taccaaaggt atcgacggcc
     3121 tgactgcgca actgaaatcg atttctcaac agaaaatcac tctggaagag aaaaaataat
     3181 gagcgagtca ctgagctgga tgcagacggg tgacacgctg gcgttatccg gagagctgga
     3241 tcaggacgtt ttgctaccgc tttgggaaat gcgtgaggaa gcggtgaagg ggattacctg
     3301 catcgatctt agccgtgtct cccgcgtgga tacgggggga ctggcactgc tgctccatct
     3361 tatcgatctg gcgaaaaagc agggcaacaa tgtgacgctt cagggggtaa acgacaaagt
     3421 gtataccctg gcgaaattgt ataatttgcc tgctgatgtt ctgcctcgtt aaggtaccaa
     3481 attccagaaa agagacgctg aaaagcgtct tttttcgttt tggtcctggt ctgaggaatg
     3541 gttgtaagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     3601 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     3661 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     3721 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3781 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3841 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     3901 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     3961 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4081 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4141 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4201 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4261 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     4321 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4381 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4441 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4501 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4561 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     4621 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4681 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4741 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4801 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4861 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4921 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct
     4981 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5041 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5101 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5161 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     5221 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5281 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     5341 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5401 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     5461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5641 ccgaaggtaa ctggctcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5701 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5761 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5821 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5881 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5941 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6001 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6061 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6121 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6181 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6241 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6301 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6361 tagggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6481 gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc
     6541 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6601 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6661 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc
     6721 ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     6781 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     6841 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     6901 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     6961 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     7021 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     7081 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     7141 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     7201 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     7261 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     7321 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     7381 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     7441 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     7501 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     7561 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     7621 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     7681 cattttgcgc ttcagccata cttttcatac tcccgc
//
