LOCUS       sequence-383002-        7674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaF-MlaE-MlaD(F118A/L123A)-MlaC-MlaB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7674)
  AUTHORS   MacRae MR, Puvanendran D, Haase MAB, Coudray N, Kolich L, Lam C, 
            Baek M, Bhabha G, Ekiert DC
  TITLE     Protein-protein interactions in the Mla lipid transport system 
            probed by computational structure prediction and deep mutational 
            scanning.
  JOURNAL   J Biol Chem. 2023 Apr 24:104744. doi: 10.1016/j.jbc.2023.104744.
  PUBMED    37100290
REFERENCE   2  (bases 1 to 7674)
  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: "J Biol 
            Chem. 2023 Apr 24:104744. doi: 10.1016/j.jbc.2023.104744."
COMMENT     Sequence Label: pBEL1953
FEATURES             Location/Qualifiers
     source          1..7674
                     /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"
     primer_bind     complement(3542..3559)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3712..3798
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3890..3917
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3936..4027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4028..4096,4097..4888)
                     /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(4246..4265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4930..5385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5017..5036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5227..5248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5496..6083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5984..6003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6237..6254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6420..6442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6580..6599
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6779..7657)
                     /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 ttaaatgcgc tggcgtcgct cggacataaa gggattaaaa ccctgagaac
     1201 gttcgggcgg gccgggttaa tgttattcaa tgcgctggtc ggcaaaccgg aatttcgcaa
     1261 acatgcgccg ctgctggtgc gccagctcta taatgtcggc gtcctgtcga tgctgattat
     1321 tgtggtttct ggcgtgttca tcggaatggt gttggggctg caaggttatc tggttctgac
     1381 cacttatagt gcggaaacca gtctgggtat gctggtggcg ttatcgctac tgcgtgaact
     1441 ggggccggtg gttgccgcgt tgttgtttgc cgggcgtgct ggttcggcgc taaccgcaga
     1501 aatcggcctg atgcgcgcta cagagcaact ctccagtatg gagatgatgg cggtggatcc
     1561 gctgcgtcgg gttatttctc cccgtttctg ggctggggtt atttcattac cactgttgac
     1621 ggttattttc gtcgccgtgg ggatctgggg cggatctctg gtcggcgtca gttggaaagg
     1681 cattgatagc gggttcttct ggtcggcaat gcaaaatgcc gtcgactggc gtatggatct
     1741 ggtcaactgt ctgattaaga gcgtggtgtt cgccatcacg gtgacgtgga tttcgttgtt
     1801 taacggctac gacgccatcc cgacgtctgc cgggattagc cgggcaacca ctcgcaccgt
     1861 tgtccactcg tctctggctg ttctggggct ggattttgtg ctgaccgcat tgatgtttgg
     1921 gaattgagtt catgcaaacg aaaaaaaatg aaatttgggt gggtatcttt ttattagcag
     1981 cactgctggc ggcgctgttt gtttgcctga aggcggcgaa cgtgacgtcc atacgtactg
     2041 aaccgaccta cacgctttat gcgacgttcg ataacattgg cggcctgaaa gcgcgctctc
     2101 cggtcagtat tggtggcgtt gttgtgggtc gggtggcgga tattacgctg gacccgaaaa
     2161 cctatctgcc gcgcgtaacg ctggaaattg aacaacgtta taaccacatt ccagatacca
     2221 gttcgctgag cattcgtact tccggcctgc tgggggaaca atatttggca ttaaacgtcg
     2281 gtgcggaaga cccggaagcg gggactgcta tcctgaagga tggcgataca attcaggaca
     2341 ctaagtctgc gatggtgctg gaagatctca ttggtcagtt cctttacggt agtaaaggcg
     2401 atgacaataa gaatagtggc gatgcgccag ctgctgcgcc aggtaataat gaaaccactg
     2461 aacctgtggg tacaacgaaa taatttcagg agaaccgacg catgtttaaa cgtttaatga
     2521 tggtcgcttt gctggtgatt gcacctctga gtgcggcaac cgcggcagac cagaccaatc
     2581 cgtataagct gatggacgag gcggcgcaga aaacgttcga tcgcctgaag aatgagcaac
     2641 cgcaaattcg ggccaacccg gattatctgc gtaccattgt tgatcaggaa ctgctgccat
     2701 acgtacaggt gaaatacgcc ggtgcgctgg tgctgggcca gtattacaag agtgcgaccc
     2761 ctgctcaacg tgaagcctac tttgccgctt tccgtgagta cctgaagcag gcttacggtc
     2821 aggcgctggc gatgtatcac ggtcaaacct atcagattgc gccagaacag ccgctgggcg
     2881 ataaaaccat tgtgcctatt cgcgttacca ttattgaccc gaatggccgt ccgccggtgc
     2941 gtctggactt ccagtggcgt aaaaactccc agacgggcaa ttggcaggct tacgacatga
     3001 ttgctgaagg cgtcagtatg atcaccacca aacaaaacga gtggggaacg ctgctgcgta
     3061 ccaaaggtat cgacggcctg actgcgcaac tgaaatcgat ttctcaacag aaaatcactc
     3121 tggaagagaa aaaataatga gcgagtcact gagctggatg cagacgggtg acacgctggc
     3181 gttatccgga gagctggatc aggacgtttt gctaccgctt tgggaaatgc gtgaggaagc
     3241 ggtgaagggg attacctgca tcgatcttag ccgtgtctcc cgcgtggata cggggggact
     3301 ggcactgctg ctccatctta tcgatctggc gaaaaagcag ggcaacaatg tgacgcttca
     3361 gggggtaaac gacaaagtgt ataccctggc gaaattgtat aatttgcctg ctgatgttct
     3421 gcctcgttaa ggtaccaaat tccagaaaag agacgctgaa aagcgtcttt tttcgttttg
     3481 gtcctggtct gaggaatggt tgtaagcttg gctgttttgg cggatgagag aagattttca
     3541 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     3601 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     3661 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     3721 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     3781 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     3841 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     3901 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     3961 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4021 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4081 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4141 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4201 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4261 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4321 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4381 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4441 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4501 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4561 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4621 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4681 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4741 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4801 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4861 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4921 gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4981 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5041 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5101 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5161 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5221 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     5281 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5341 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg
     5401 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5461 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5521 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5581 gagctaccaa ctctttttcc gaaggtaact ggctcagcag agcgcagata ccaaatactg
     5641 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5701 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5761 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5821 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5881 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5941 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6001 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6061 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6121 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6181 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6241 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     6301 gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa cacccgctga
     6361 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6421 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga
     6481 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     6541 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     6601 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     6661 ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag
     6721 ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt
     6781 atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg
     6841 ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa
     6901 cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc
     6961 tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat
     7021 gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc
     7081 tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg
     7141 gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta
     7201 tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca
     7261 ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata
     7321 ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt
     7381 gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca
     7441 gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac
     7501 cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat
     7561 cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt
     7621 atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc ccgc
//
