LOCUS       sequence-320082-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for MlaE Y81W.
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: pBEL2100
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..329
                     /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     252..271
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1970..1987
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2000..2020
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3631..3648)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3801..3887
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3979..4006
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4025..4116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4117..4185,4186..4977)
                     /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(4335..4354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5019..5474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5106..5125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5316..5337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5585..6172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6073..6092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6326..6343
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6509..6531
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6669..6688
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6868..7728,1..18))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
       61 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
      121 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
      181 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
      241 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
      301 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagcc gaagggtgaa
      361 ttatggagca gtctgtggcg aatttagtcg atatgcgcga tgtcagtttt acgcgtggca
      421 atcgctgcat cttcgataat atttccctga ccgtgccgcg agggaagatc acggcgatca
      481 tggggccatc gggcatcggt aaaacgacgc tactccgtct gattggcggg caaatcgcac
      541 cagatcatgg tgagatcctt ttcgatggtg agaatattcc ggcgatgtct cgttcgcgcc
      601 tgtatacagt gcgcaaacgg atgagcatgt tatttcagtc cggggcgttg ttcactgata
      661 tgaacgtatt tgacaacgtc gcctatccac tgcgcgaaca tacccaactt cccgcgccat
      721 tgttgcatag tacggtgatg atgaagctgg aggccgtggg gctgcgtgga gcggctaaac
      781 taatgccttc tgaactttcc ggtgggatgg cgcggcgtgc agcgctggcg cgtgcgattg
      841 cgctggagcc ggatctcatc atgtttgatg aaccttttgt tgggcaagat cccatcacca
      901 tgggcgtact ggtgaagctg atttctgagc tgaacagcgc gctgggcgtg acttgtgtgg
      961 tggtttctca cgatgtgccg gaagtgttaa gtattgcgga tcacgcctgg atcctggcgg
     1021 acaaaaaaat tgtcgctcat ggcagtgccc aggcgttgca ggcgaatcct gatccgcgcg
     1081 tacgtcagtt tctggacggg atagctgacg ggcctgttcc gttccgctat cctgccggcg
     1141 attatcacgc tgatctttta ccagggagtt aagccactca tgctgttaaa tgcgctggcg
     1201 tcgctcggac ataaagggat taaaaccctg agaacgttcg ggcgggccgg gttaatgtta
     1261 ttcaatgcgc tggtcggcaa accggaattt cgcaaacatg cgccgctgct ggtgcgccag
     1321 ctctataatg tcggcgtcct gtcgatgctg attattgtgg tttctggcgt gttcatcgga
     1381 atggtgttgg ggctgcaagg ttatctggtt ctgaccactt ggagtgcgga aaccagtctg
     1441 ggtatgctgg tggcgttatc gctactgcgt gaactggggc cggtggttgc cgcgttgttg
     1501 tttgccgggc gtgctggttc ggcgctaacc gcagaaatcg gcctgatgcg cgctacagag
     1561 caactctcca gtatggagat gatggcggtg gatccgctgc gtcgggttat ttctccccgt
     1621 ttctgggctg gggttatttc attaccactg ttgacggtta ttttcgtcgc cgtggggatc
     1681 tggggcggat ctctggtcgg cgtcagttgg aaaggcattg atagcgggtt cttctggtcg
     1741 gcaatgcaaa atgccgtcga ctggcgtatg gatctggtca actgtctgat taagagcgtg
     1801 gtgttcgcca tcacggtgac gtggatttcg ttgtttaacg gctacgacgc catcccgacg
     1861 tctgccggga ttagccgggc aaccactcgc accgttgtcc actcgtctct ggctgttctg
     1921 gggctggatt ttgtgctgac cgcattgatg tttgggaatt gagttcatgc atcaccacca
     1981 tcaccatcag catcaacacg aaaatctgta ttttcagggg atgcaaacga aaaaaaatga
     2041 aatttgggtg ggtatctttt tattagcagc actgctggcg gcgctgtttg tttgcctgaa
     2101 ggcggcgaac gtgacgtcca tacgtactga accgacctac acgctttatg cgacgttcga
     2161 taacattggc ggcctgaaag cgcgctctcc ggtcagtatt ggtggcgttg ttgtgggtcg
     2221 ggtggcggat attacgctgg acccgaaaac ctatctgccg cgcgtaacgc tggaaattga
     2281 acaacgttat aaccacattc cagataccag ttcgctgagc attcgtactt ccggcctgct
     2341 gggggaacaa tatctggcat taaacgtcgg ttttgaagac ccggaactgg ggactgctat
     2401 cctgaaggat ggcgatacaa ttcaggacac taagtctgcg atggtgctgg aagatctcat
     2461 tggtcagttc ctttacggta gtaaaggcga tgacaataag aatagtggcg atgcgccagc
     2521 tgctgcgcca ggtaataatg aaaccactga acctgtgggt acaacgaaat aatttcagga
     2581 gaaccgacgc atgtttaaac gtttaatgat ggtcgctttg ctggtgattg cacctctgag
     2641 tgcggcaacc gcggcagacc agaccaatcc gtataagctg atggacgagg cggcgcagaa
     2701 aacgttcgat cgcctgaaga atgagcaacc gcaaattcgg gccaacccgg attatctgcg
     2761 taccattgtt gatcaggaac tgctgccata cgtacaggtg aaatacgccg gtgcgctggt
     2821 gctgggccag tattacaaga gtgcgacccc tgctcaacgt gaagcctact ttgccgcttt
     2881 ccgtgagtac ctgaagcagg cttacggtca ggcgctggcg atgtatcacg gtcaaaccta
     2941 tcagattgcg ccagaacagc cgctgggcga taaaaccatt gtgcctattc gcgttaccat
     3001 tattgacccg aatggccgtc cgccggtgcg tctggacttc cagtggcgta aaaactccca
     3061 gacgggcaat tggcaggctt acgacatgat tgctgaaggc gtcagtatga tcaccaccaa
     3121 acaaaacgag tggggaacgc tgctgcgtac caaaggtatc gacggcctga ctgcgcaact
     3181 gaaatcgatt tctcaacaga aaatcactct ggaagagaaa aaataatgag cgagtcactg
     3241 agctggatgc agacgggtga cacgctggcg ttatccggag agctggatca ggacgttttg
     3301 ctaccgcttt gggaaatgcg tgaggaagcg gtgaagggga ttacctgcat cgatcttagc
     3361 cgtgtctccc gcgtggatac ggggggactg gcactgctgc tccatcttat cgatctggcg
     3421 aaaaagcagg gcaacaatgt gacgcttcag ggggtaaacg acaaagtgta taccctggcg
     3481 aaattgtata atttgcctgc tgatgttctg cctcgttaag gtaccaaatt ccagaaaaga
     3541 gacgctgaaa agcgtctttt ttcgttttgg tcctggtctg aggaatggtt gtaagcttgg
     3601 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     3661 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     3721 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     3781 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3841 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     3901 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     3961 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     4021 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4081 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4141 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4201 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4261 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4321 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4381 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4501 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4621 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     4681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4981 tcagaccaag tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt
     5041 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5101 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5161 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5221 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5281 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5341 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     5401 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5461 aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5521 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5581 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5641 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5701 gctcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5761 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5821 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5881 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5941 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6001 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6061 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6121 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6181 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6241 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6301 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     6361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag ggtcatggct
     6421 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6481 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6541 tcatcaccga aacgcgcgag gcagaaggag atggcgccca acagtccccc ggccacgggg
     6601 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     6661 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     6721 ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tcatcgatgc
     6781 ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa
     6841 gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt
     6901 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     6961 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     7021 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     7081 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     7141 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     7201 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     7261 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     7321 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     7381 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     7441 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     7501 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     7561 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     7621 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     7681 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatc
//
