LOCUS       sequence-382999-        5233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaC(R147D).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5233)
  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 5233)
  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: pBEL2536
FEATURES             Location/Qualifiers
     source          1..5233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     277..296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     530..547
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     713..735
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     873..892
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1072..1950)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1977..2261
                     /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     2184..2203
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3068..3085)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3238..3324
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3416..3443
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3462..3553
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3554..3622,3623..4414)
                     /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(3772..3791)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4456..4911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4543..4562)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4753..4774
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      join(5022..5233,1..376)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
       61 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      421 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      481 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      541 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
      601 ttcacaccgc atagggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
      661 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
      721 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg
      781 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
      841 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      901 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca
      961 tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct
     1021 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     1081 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     1141 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     1201 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     1261 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     1321 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     1381 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     1441 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     1501 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     1561 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     1621 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     1681 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     1741 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     1801 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     1861 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     1921 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     1981 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     2041 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     2101 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     2161 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     2221 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag
     2281 cggccggcct ggcccgaagg gtgaattatg tttaaacgtt taatgatggt cgctttgctg
     2341 gtgattgcac ctctgagtgc ggcaaccgcg gcagaccaga ccaatccgta taagctgatg
     2401 gacgaggcgg cgcagaaaac gttcgatcgc ctgaagaatg agcaaccgca aattcgggcc
     2461 aacccggatt atctgcgtac cattgttgat caggaactgc tgccatacgt acaggtgaaa
     2521 tacgccggtg cgctggtgct gggccagtat tacaagagtg cgacccctgc tcaacgtgaa
     2581 gcctactttg ccgctttccg tgagtacctg aagcaggctt acggtcaggc gctggcgatg
     2641 tatcacggtc aaacctatca gattgcgcca gaacagccgc tgggcgataa aaccattgtg
     2701 cctattcgcg ttaccattat tgacccgaat ggccgtccgc cggtggatct ggacttccag
     2761 tggcgtaaaa actcccagac gggcaattgg caggcttacg acatgattgc tgaaggcgtc
     2821 agtatgatca ccaccaaaca aaacgagtgg ggaacgctgc tgcgtaccaa aggtatcgac
     2881 ggcctgactg cgcaactgaa atcgatttct caacagaaaa tcactctgga agagaaaaaa
     2941 taaggcccag gcggccggta ccaaattcca gaaaagagac gctgaaaagc gtcttttttc
     3001 gttttggtcc tggtctgagg aatggttgta agcttggctg ttttggcgga tgagagaaga
     3061 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     3121 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     3181 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     3241 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3301 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     3361 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     3421 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     3481 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3541 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3601 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3661 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3721 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3781 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     3841 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3901 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3961 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4021 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4081 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4141 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4201 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4261 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4321 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4381 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4441 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4501 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4561 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4621 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4681 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4741 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4801 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4861 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     4921 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5101 gatcaagagc taccaactct ttttccgaag gtaactggct cagcagagcg cagataccaa
     5161 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5221 ctacatacct cgc
//
