LOCUS       sequence-385337-        7581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaF-MlaE-MlaDdelta153-183-MlaC-MlaB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  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 7581)
  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: pBEL1936
FEATURES             Location/Qualifiers
     source          1..7581
                     /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(3449..3466)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3619..3705
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3797..3824
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3843..3934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3935..4003,4004..4795)
                     /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(4153..4172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4837..5292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4924..4943)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5134..5155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5403..5990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5891..5910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6144..6161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6327..6349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6487..6506
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6686..7564)
                     /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 atatctggca ttaaacgtcg
     2281 gttttgaaga cccggaactg gggactgcta tcctgaagga tggcgataca attcaggaca
     2341 ctaagtctgc gatggtgctg gaagatctca ttggtcagtt cctttactaa tttcaggaga
     2401 accgacgcat gtttaaacgt ttaatgatgg tcgctttgct ggtgattgca cctctgagtg
     2461 cggcaaccgc ggcagaccag accaatccgt ataagctgat ggacgaggcg gcgcagaaaa
     2521 cgttcgatcg cctgaagaat gagcaaccgc aaattcgggc caacccggat tatctgcgta
     2581 ccattgttga tcaggaactg ctgccatacg tacaggtgaa atacgccggt gcgctggtgc
     2641 tgggccagta ttacaagagt gcgacccctg ctcaacgtga agcctacttt gccgctttcc
     2701 gtgagtacct gaagcaggct tacggtcagg cgctggcgat gtatcacggt caaacctatc
     2761 agattgcgcc agaacagccg ctgggcgata aaaccattgt gcctattcgc gttaccatta
     2821 ttgacccgaa tggccgtccg ccggtgcgtc tggacttcca gtggcgtaaa aactcccaga
     2881 cgggcaattg gcaggcttac gacatgattg ctgaaggcgt cagtatgatc accaccaaac
     2941 aaaacgagtg gggaacgctg ctgcgtacca aaggtatcga cggcctgact gcgcaactga
     3001 aatcgatttc tcaacagaaa atcactctgg aagagaaaaa ataatgagcg agtcactgag
     3061 ctggatgcag acgggtgaca cgctggcgtt atccggagag ctggatcagg acgttttgct
     3121 accgctttgg gaaatgcgtg aggaagcggt gaaggggatt acctgcatcg atcttagccg
     3181 tgtctcccgc gtggatacgg ggggactggc actgctgctc catcttatcg atctggcgaa
     3241 aaagcagggc aacaatgtga cgcttcaggg ggtaaacgac aaagtgtata ccctggcgaa
     3301 attgtataat ttgcctgctg atgttctgcc tcgttaaggt accaaattcc agaaaagaga
     3361 cgctgaaaag cgtctttttt cgttttggtc ctggtctgag gaatggttgt aagcttggct
     3421 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     3481 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     3541 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     3601 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3661 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     3721 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     3781 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     3841 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4441 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4801 agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc ggcgcattaa
     4861 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4921 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4981 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5041 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5101 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5161 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     5221 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5281 cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5341 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5401 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5461 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5521 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5581 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5641 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5701 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5761 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5821 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5881 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5941 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6001 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6061 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6121 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     6181 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg tcatggctgc
     6241 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6301 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6361 atcaccgaaa cgcgcgaggc agaaggagat ggcgcccaac agtcccccgg ccacggggcc
     6421 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     6481 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6541 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc atcgatgcat
     6601 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     6661 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     6721 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     6781 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     6841 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     6901 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     6961 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     7021 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     7081 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     7141 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     7201 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     7261 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     7321 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     7381 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     7441 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     7501 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     7561 ccatactttt catactcccg c
//
