LOCUS       sequence-383742-        5314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaA(F223N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5314)
  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 5314)
  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: pBEL2663
FEATURES             Location/Qualifiers
     source          1..5314
                     /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(1182..1199)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1352..1438
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1530..1557
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1576..1667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1668..1736,1737..2528)
                     /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(1886..1905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2570..3025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2657..2676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2867..2888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3136..3723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3624..3643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3877..3894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4060..4082
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4220..4239
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4419..5297)
                     /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 tagcatgaag cttcgcctgt cggcgcttgc tctgggaact acgcttctgg
      361 tggggtgtgc gagttccggt acagatcagc aagggcgttc tgacccgtta gaagggttca
      421 accgcaccat gtacaacttc aacttcaatg tattagaccc gtatattgtt cgaccggtcg
      481 ctgtcgcctg gcgtgattat gttccgcaac cggcgcgtaa cggtttgagc aactttactg
      541 gcaaccttga agaacctgcg gtgatggtta actacttctt gcagggcgac ccttatcagg
      601 ggatggtcca ctttacccgc tttttcctga acaccatttt ggggatgggc ggttttattg
      661 atgttgcagg gatggcgaac ccgaaactgc aacggactga acctcaccgc ttcggtagta
      721 cgcttggtca ttatggcgtg ggttatgggc cttacgttca gttaccgttc tacggtagct
      781 tcacgctgcg tgatgacggt ggtgatatgg cggatggttt ttacccggtt ctttcctggc
      841 tgacctggcc gatgtctgtg ggtaaatgga cgcttgaagg gatcgaaacc cgcgctcagc
      901 tgctggattc cgatggtctg ctgcgtcagt cgtccgatcc ttatattatg gtgcgcgaag
      961 cgtacttcca gcgtcatgat aacatcgcta atggcggcga actcaaaccg caggaaaatc
     1021 cgaacgcaca agcgattcag gatgatttaa aagatattga ttctgaataa ggtaccaaat
     1081 tccagaaaag agacgctgaa aagcgtcttt tttcgttttg gtcctggtct gaggaatggt
     1141 tgtaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     1201 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     1261 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     1321 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1381 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1441 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     1501 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     1561 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     1921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2041 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     2221 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2281 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2521 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt
     2581 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2641 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2701 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2761 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2821 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2881 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     2941 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3001 aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct
     3061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3241 gaaggtaact ggctcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3781 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3841 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3901 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3961 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4021 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4081 ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc
     4141 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     4201 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4261 cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
     4321 atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
     4381 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
     4441 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     4501 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     4561 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     4621 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     4681 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     4741 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     4801 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     4861 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     4921 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     4981 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     5041 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     5101 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     5161 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     5221 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     5281 ttttgcgctt cagccatact tttcatactc ccgc
//
