LOCUS       sequence-342022-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and complementation plasmid for MlaFEDCB, with N-terminal
            his tag on MlaE.
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: pBEL1198
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..366
                     /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     289..308
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1220..1237
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1250..1270
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3668..3685)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3838..3924
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4016..4043
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4062..4153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4154..4222,4223..5014)
                     /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(4372..4391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5056..5511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5143..5162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5353..5374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5622..6209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6110..6129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6363..6380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6546..6568
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6706..6725
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6905..7728,1..55))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
       61 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
      121 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
      181 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
      241 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
      301 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
      361 ctccataccc gtttttttgg gctagccgaa gggtgaatta tggagcagtc tgtggcgaat
      421 ttagtcgata tgcgcgatgt cagttttacg cgtggcaatc gctgcatctt cgataatatt
      481 tccctgaccg tgccgcgagg gaagatcacg gcgatcatgg ggccatcggg catcggtaaa
      541 acgacgctac tccgtctgat tggcgggcaa atcgcaccag atcatggtga gatccttttc
      601 gatggtgaga atattccggc gatgtctcgt tcgcgcctgt atacagtgcg caaacggatg
      661 agcatgttat ttcagtccgg ggcgttgttc actgatatga acgtatttga caacgtcgcc
      721 tatccactgc gcgaacatac ccaacttccc gcgccattgt tgcatagtac ggtgatgatg
      781 aagctggagg ccgtggggct gcgtggagcg gctaaactaa tgccttctga actttccggt
      841 gggatggcgc ggcgtgcagc gctggcgcgt gcgattgcgc tggagccgga tctcatcatg
      901 tttgatgaac cttttgttgg gcaagatccc atcaccatgg gcgtactggt gaagctgatt
      961 tctgagctga acagcgcgct gggcgtgact tgtgtggtgg tttctcacga tgtgccggaa
     1021 gtgttaagta ttgcggatca cgcctggatc ctggcggaca aaaaaattgt cgctcatggc
     1081 agtgcccagg cgttgcaggc gaatcctgat ccgcgcgtac gtcagtttct ggacgggata
     1141 gctgacgggc ctgttccgtt ccgctatcct gccggcgatt atcacgctga tcttttacca
     1201 gggagttaag ccactcatgc atcaccacca tcaccatcag catcaacacg aaaatctgta
     1261 ttttcagggg atgctgttaa atgcgctggc gtcgctcgga cataaaggga ttaaaaccct
     1321 gagaacgttc gggcgggccg ggttaatgtt attcaatgcg ctggtcggca aaccggaatt
     1381 tcgcaaacat gcgccgctgc tggtgcgcca gctctataat gtcggcgtcc tgtcgatgct
     1441 gattattgtg gtttctggcg tgttcatcgg aatggtgttg gggctgcaag gttatctggt
     1501 tctgaccact tatagtgcgg aaaccagtct gggtatgctg gtggcgttat cgctactgcg
     1561 tgaactgggg ccggtggttg ccgcgttgtt gtttgccggg cgtgctggtt cggcgctaac
     1621 cgcagaaatc ggcctgatgc gcgctacaga gcaactctcc agtatggaga tgatggcggt
     1681 ggatccgctg cgtcgggtta tttctccccg tttctgggct ggggttattt cattaccact
     1741 gttgacggtt attttcgtcg ccgtggggat ctggggcgga tctctggtcg gcgtcagttg
     1801 gaaaggcatt gatagcgggt tcttctggtc ggcaatgcaa aatgccgtcg actggcgtat
     1861 ggatctggtc aactgtctga ttaagagcgt ggtgttcgcc atcacggtga cgtggatttc
     1921 gttgtttaac ggctacgacg ccatcccgac gtctgccggg attagccggg caaccactcg
     1981 caccgttgtc cactcgtctc tggctgttct ggggctggat tttgtgctga ccgcattgat
     2041 gtttgggaat tgagttcatg caaacgaaaa aaaatgaaat ttgggtgggt atctttttat
     2101 tagcagcact gctggcggcg ctgtttgttt gcctgaaggc ggcgaacgtg acgtccatac
     2161 gtactgaacc gacctacacg ctttatgcga cgttcgataa cattggcggc ctgaaagcgc
     2221 gctctccggt cagtattggt ggcgttgttg tgggtcgggt ggcggatatt acgctggacc
     2281 cgaaaaccta tctgccgcgc gtaacgctgg aaattgaaca acgttataac cacattccag
     2341 ataccagttc gctgagcatt cgtacttccg gcctgctggg ggaacaatat ctggcattaa
     2401 acgtcggttt tgaagacccg gaactgggga ctgctatcct gaaggatggc gatacaattc
     2461 aggacactaa gtctgcgatg gtgctggaag atctcattgg tcagttcctt tacggtagta
     2521 aaggcgatga caataagaat agtggcgatg cgccagctgc tgcgccaggt aataatgaaa
     2581 ccactgaacc tgtgggtaca acgaaataat ttcaggagaa ccgacgcatg tttaaacgtt
     2641 taatgatggt cgctttgctg gtgattgcac ctctgagtgc ggcaaccgcg gcagaccaga
     2701 ccaatccgta taagctgatg gacgaggcgg cgcagaaaac gttcgatcgc ctgaagaatg
     2761 agcaaccgca aattcgggcc aacccggatt atctgcgtac cattgttgat caggaactgc
     2821 tgccatacgt acaggtgaaa tacgccggtg cgctggtgct gggccagtat tacaagagtg
     2881 cgacccctgc tcaacgtgaa gcctactttg ccgctttccg tgagtacctg aagcaggctt
     2941 acggtcaggc gctggcgatg tatcacggtc aaacctatca gattgcgcca gaacagccgc
     3001 tgggcgataa aaccattgtg cctattcgcg ttaccattat tgacccgaat ggccgtccgc
     3061 cggtgcgtct ggacttccag tggcgtaaaa actcccagac gggcaattgg caggcttacg
     3121 acatgattgc tgaaggcgtc agtatgatca ccaccaaaca aaacgagtgg ggaacgctgc
     3181 tgcgtaccaa aggtatcgac ggcctgactg cgcaactgaa atcgatttct caacagaaaa
     3241 tcactctgga agagaaaaaa taatgagcga gtcactgagc tggatgcaga cgggtgacac
     3301 gctggcgtta tccggagagc tggatcagga cgttttgcta ccgctttggg aaatgcgtga
     3361 ggaagcggtg aaggggatta cctgcatcga tcttagccgt gtctcccgcg tggatacggg
     3421 gggactggca ctgctgctcc atcttatcga tctggcgaaa aagcagggca acaatgtgac
     3481 gcttcagggg gtaaacgaca aagtgtatac cctggcgaaa ttgtataatt tgcctgctga
     3541 tgttctgcct cgttaaggta ccaaattcca gaaaagagac gctgaaaagc gtcttttttc
     3601 gttttggtcc tggtctgagg aatggttgta agcttggctg ttttggcgga tgagagaaga
     3661 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     3721 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     3781 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     3841 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3901 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     3961 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     4021 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     4081 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4141 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4201 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4261 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4321 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4381 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     4441 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4501 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4561 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4621 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4681 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4741 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4801 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4861 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4921 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4981 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5041 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5101 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5161 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5221 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5281 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5341 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     5401 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5461 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     5521 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5581 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5641 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5701 gatcaagagc taccaactct ttttccgaag gtaactggct cagcagagcg cagataccaa
     5761 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6241 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6301 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6361 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     6421 atctgtgcgg tatttcacac cgcatagggt catggctgcg ccccgacacc cgccaacacc
     6481 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6541 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     6601 gaaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     6661 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     6721 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     6781 gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg
     6841 aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta
     6901 ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
     6961 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
     7021 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
     7081 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
     7141 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
     7201 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
     7261 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
     7321 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
     7381 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
     7441 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     7501 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     7561 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     7621 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     7681 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccacca
//
