LOCUS       sequence-320154-        7641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaE IF1 1-30 aa 
            deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7641)
  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 7641)
  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: pBEL2092
FEATURES             Location/Qualifiers
     source          1..7641
                     /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             1848..1865
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1878..1898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3509..3526)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3679..3765
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3857..3884
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3903..3994
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3995..4063,4064..4855)
                     /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(4213..4232)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4897..5352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4984..5003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5194..5215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5463..6050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5951..5970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6204..6221
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6387..6409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6547..6566
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6746..7624)
                     /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 actcatgctg gtcggcaaac cggaatttcg caaacatgcg ccgctgctgg tgcgccagct
     1201 ctataatgtc ggcgtcctgt cgatgctgat tattgtggtt tctggcgtgt tcatcggaat
     1261 ggtgttgggg ctgcaaggtt atctggttct gaccacttat agtgcggaaa ccagtctggg
     1321 tatgctggtg gcgttatcgc tactgcgtga actggggccg gtggttgccg cgttgttgtt
     1381 tgccgggcgt gctggttcgg cgctaaccgc agaaatcggc ctgatgcgcg ctacagagca
     1441 actctccagt atggagatga tggcggtgga tccgctgcgt cgggttattt ctccccgttt
     1501 ctgggctggg gttatttcat taccactgtt gacggttatt ttcgtcgccg tggggatctg
     1561 gggcggatct ctggtcggcg tcagttggaa aggcattgat agcgggttct tctggtcggc
     1621 aatgcaaaat gccgtcgact ggcgtatgga tctggtcaac tgtctgatta agagcgtggt
     1681 gttcgccatc acggtgacgt ggatttcgtt gtttaacggc tacgacgcca tcccgacgtc
     1741 tgccgggatt agccgggcaa ccactcgcac cgttgtccac tcgtctctgg ctgttctggg
     1801 gctggatttt gtgctgaccg cattgatgtt tgggaattga gttcatgcat caccaccatc
     1861 accatcagca tcaacacgaa aatctgtatt ttcaggggat gcaaacgaaa aaaaatgaaa
     1921 tttgggtggg tatcttttta ttagcagcac tgctggcggc gctgtttgtt tgcctgaagg
     1981 cggcgaacgt gacgtccata cgtactgaac cgacctacac gctttatgcg acgttcgata
     2041 acattggcgg cctgaaagcg cgctctccgg tcagtattgg tggcgttgtt gtgggtcggg
     2101 tggcggatat tacgctggac ccgaaaacct atctgccgcg cgtaacgctg gaaattgaac
     2161 aacgttataa ccacattcca gataccagtt cgctgagcat tcgtacttcc ggcctgctgg
     2221 gggaacaata tctggcatta aacgtcggtt ttgaagaccc ggaactgggg actgctatcc
     2281 tgaaggatgg cgatacaatt caggacacta agtctgcgat ggtgctggaa gatctcattg
     2341 gtcagttcct ttacggtagt aaaggcgatg acaataagaa tagtggcgat gcgccagctg
     2401 ctgcgccagg taataatgaa accactgaac ctgtgggtac aacgaaataa tttcaggaga
     2461 accgacgcat gtttaaacgt ttaatgatgg tcgctttgct ggtgattgca cctctgagtg
     2521 cggcaaccgc ggcagaccag accaatccgt ataagctgat ggacgaggcg gcgcagaaaa
     2581 cgttcgatcg cctgaagaat gagcaaccgc aaattcgggc caacccggat tatctgcgta
     2641 ccattgttga tcaggaactg ctgccatacg tacaggtgaa atacgccggt gcgctggtgc
     2701 tgggccagta ttacaagagt gcgacccctg ctcaacgtga agcctacttt gccgctttcc
     2761 gtgagtacct gaagcaggct tacggtcagg cgctggcgat gtatcacggt caaacctatc
     2821 agattgcgcc agaacagccg ctgggcgata aaaccattgt gcctattcgc gttaccatta
     2881 ttgacccgaa tggccgtccg ccggtgcgtc tggacttcca gtggcgtaaa aactcccaga
     2941 cgggcaattg gcaggcttac gacatgattg ctgaaggcgt cagtatgatc accaccaaac
     3001 aaaacgagtg gggaacgctg ctgcgtacca aaggtatcga cggcctgact gcgcaactga
     3061 aatcgatttc tcaacagaaa atcactctgg aagagaaaaa ataatgagcg agtcactgag
     3121 ctggatgcag acgggtgaca cgctggcgtt atccggagag ctggatcagg acgttttgct
     3181 accgctttgg gaaatgcgtg aggaagcggt gaaggggatt acctgcatcg atcttagccg
     3241 tgtctcccgc gtggatacgg ggggactggc actgctgctc catcttatcg atctggcgaa
     3301 aaagcagggc aacaatgtga cgcttcaggg ggtaaacgac aaagtgtata ccctggcgaa
     3361 attgtataat ttgcctgctg atgttctgcc tcgttaaggt accaaattcc agaaaagaga
     3421 cgctgaaaag cgtctttttt cgttttggtc ctggtctgag gaatggttgt aagcttggct
     3481 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     3541 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     3601 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     3661 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3721 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     3781 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     3841 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     3901 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3961 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4021 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4081 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4141 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4201 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4261 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4321 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4381 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4441 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4501 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     4561 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4621 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4681 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4741 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4801 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4861 agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc ggcgcattaa
     4921 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4981 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5041 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5101 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5161 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5221 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     5281 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5341 cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5401 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5461 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5521 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5581 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5821 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6121 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6181 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     6241 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg tcatggctgc
     6301 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6361 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6421 atcaccgaaa cgcgcgaggc agaaggagat ggcgcccaac agtcccccgg ccacggggcc
     6481 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     6541 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6601 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc atcgatgcat
     6661 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     6721 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     6781 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     6841 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     6901 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     6961 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     7021 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     7081 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     7141 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     7201 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     7261 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     7321 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     7381 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     7441 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     7501 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     7561 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     7621 ccatactttt catactcccg c
//
