LOCUS       sequence-385689-        7674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaF-MlaE-MlaD-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: pBEL1195
FEATURES             Location/Qualifiers
     source          1..7674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..293
                     /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     216..235
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3541..3558)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3711..3797
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3889..3916
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3935..4026
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4027..4095,4096..4887)
                     /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(4245..4264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4929..5384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5016..5035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5226..5247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5495..6082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5983..6002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6236..6253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6419..6441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6579..6598
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6778..7656)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
       61 ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag
      121 cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt
      181 ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag
      241 attagcggat cctacctgac gctttttatc gcaactctct actgtttctc catacccgtt
      301 tttttgggct agccgaaggg tgaattatgg agcagtctgt ggcgaattta gtcgatatgc
      361 gcgatgtcag ttttacgcgt ggcaatcgct gcatcttcga taatatttcc ctgaccgtgc
      421 cgcgagggaa gatcacggcg atcatggggc catcgggcat cggtaaaacg acgctactcc
      481 gtctgattgg cgggcaaatc gcaccagatc atggtgagat ccttttcgat ggtgagaata
      541 ttccggcgat gtctcgttcg cgcctgtata cagtgcgcaa acggatgagc atgttatttc
      601 agtccggggc gttgttcact gatatgaacg tatttgacaa cgtcgcctat ccactgcgcg
      661 aacataccca acttcccgcg ccattgttgc atagtacggt gatgatgaag ctggaggccg
      721 tggggctgcg tggagcggct aaactaatgc cttctgaact ttccggtggg atggcgcggc
      781 gtgcagcgct ggcgcgtgcg attgcgctgg agccggatct catcatgttt gatgaacctt
      841 ttgttgggca agatcccatc accatgggcg tactggtgaa gctgatttct gagctgaaca
      901 gcgcgctggg cgtgacttgt gtggtggttt ctcacgatgt gccggaagtg ttaagtattg
      961 cggatcacgc ctggatcctg gcggacaaaa aaattgtcgc tcatggcagt gcccaggcgt
     1021 tgcaggcgaa tcctgatccg cgcgtacgtc agtttctgga cgggatagct gacgggcctg
     1081 ttccgttccg ctatcctgcc ggcgattatc acgctgatct tttaccaggg agttaagcca
     1141 ctcatgctgt taaatgcgct ggcgtcgctc ggacataaag ggattaaaac cctgagaacg
     1201 ttcgggcggg ccgggttaat gttattcaat gcgctggtcg gcaaaccgga atttcgcaaa
     1261 catgcgccgc tgctggtgcg ccagctctat aatgtcggcg tcctgtcgat gctgattatt
     1321 gtggtttctg gcgtgttcat cggaatggtg ttggggctgc aaggttatct ggttctgacc
     1381 acttatagtg cggaaaccag tctgggtatg ctggtggcgt tatcgctact gcgtgaactg
     1441 gggccggtgg ttgccgcgtt gttgtttgcc gggcgtgctg gttcggcgct aaccgcagaa
     1501 atcggcctga tgcgcgctac agagcaactc tccagtatgg agatgatggc ggtggatccg
     1561 ctgcgtcggg ttatttctcc ccgtttctgg gctggggtta tttcattacc actgttgacg
     1621 gttattttcg tcgccgtggg gatctggggc ggatctctgg tcggcgtcag ttggaaaggc
     1681 attgatagcg ggttcttctg gtcggcaatg caaaatgccg tcgactggcg tatggatctg
     1741 gtcaactgtc tgattaagag cgtggtgttc gccatcacgg tgacgtggat ttcgttgttt
     1801 aacggctacg acgccatccc gacgtctgcc gggattagcc gggcaaccac tcgcaccgtt
     1861 gtccactcgt ctctggctgt tctggggctg gattttgtgc tgaccgcatt gatgtttggg
     1921 aattgagttc atgcaaacga aaaaaaatga aatttgggtg ggtatctttt tattagcagc
     1981 actgctggcg gcgctgtttg tttgcctgaa ggcggcgaac gtgacgtcca tacgtactga
     2041 accgacctac acgctttatg cgacgttcga taacattggc ggcctgaaag cgcgctctcc
     2101 ggtcagtatt ggtggcgttg ttgtgggtcg ggtggcggat attacgctgg acccgaaaac
     2161 ctatctgccg cgcgtaacgc tggaaattga acaacgttat aaccacattc cagataccag
     2221 ttcgctgagc attcgtactt ccggcctgct gggggaacaa tatctggcat taaacgtcgg
     2281 ttttgaagac ccggaactgg ggactgctat cctgaaggat ggcgatacaa ttcaggacac
     2341 taagtctgcg atggtgctgg aagatctcat tggtcagttc ctttacggta gtaaaggcga
     2401 tgacaataag aatagtggcg atgcgccagc tgctgcgcca ggtaataatg aaaccactga
     2461 acctgtgggt acaacgaaat aatttcagga gaaccgacgc atgtttaaac gtttaatgat
     2521 ggtcgctttg ctggtgattg cacctctgag tgcggcaacc gcggcagacc agaccaatcc
     2581 gtataagctg atggacgagg cggcgcagaa aacgttcgat cgcctgaaga atgagcaacc
     2641 gcaaattcgg gccaacccgg attatctgcg taccattgtt gatcaggaac tgctgccata
     2701 cgtacaggtg aaatacgccg gtgcgctggt gctgggccag tattacaaga gtgcgacccc
     2761 tgctcaacgt gaagcctact ttgccgcttt ccgtgagtac ctgaagcagg cttacggtca
     2821 ggcgctggcg atgtatcacg gtcaaaccta tcagattgcg ccagaacagc cgctgggcga
     2881 taaaaccatt gtgcctattc gcgttaccat tattgacccg aatggccgtc cgccggtgcg
     2941 tctggacttc cagtggcgta aaaactccca gacgggcaat tggcaggctt acgacatgat
     3001 tgctgaaggc gtcagtatga tcaccaccaa acaaaacgag tggggaacgc tgctgcgtac
     3061 caaaggtatc gacggcctga ctgcgcaact gaaatcgatt tctcaacaga aaatcactct
     3121 ggaagagaaa aaataatgag cgagtcactg agctggatgc agacgggtga cacgctggcg
     3181 ttatccggag agctggatca ggacgttttg ctaccgcttt gggaaatgcg tgaggaagcg
     3241 gtgaagggga ttacctgcat cgatcttagc cgtgtctccc gcgtggatac ggggggactg
     3301 gcactgctgc tccatcttat cgatctggcg aaaaagcagg gcaacaatgt gacgcttcag
     3361 ggggtaaacg acaaagtgta taccctggcg aaattgtata atttgcctgc tgatgttctg
     3421 cctcgttaag gtaccaaatt ccagaaaaga gacgctgaaa agcgtctttt ttcgttttgg
     3481 tcctggtctg aggaatggtt gtaagcttgg ctgttttggc ggatgagaga agattttcag
     3541 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     3601 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     3661 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     3721 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     3781 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     3841 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     3901 tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca
     3961 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4021 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4081 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4141 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4201 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4261 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4321 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4381 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4441 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4501 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4561 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4621 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4681 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4741 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4801 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4861 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4921 attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4981 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5041 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5101 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     5161 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5221 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     5281 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5341 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga
     5401 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5461 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5521 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5581 agctaccaac tctttttccg aaggtaactg gctcagcaga gcgcagatac caaatactgt
     5641 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6181 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6241 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     6301 cggtatttca caccgcatag ggtcatggct gcgccccgac acccgccaac acccgctgac
     6361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6421 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagaaggag
     6481 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     6541 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     6601 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     6661 tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg
     6721 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
     6781 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
     6841 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
     6901 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
     6961 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
     7021 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
     7081 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
     7141 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
     7201 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     7261 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     7321 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     7381 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     7441 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     7501 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     7561 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     7621 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgcc
//
