LOCUS       sequence-383006-        5290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaA(delta244-251).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5290)
  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 5290)
  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: pBEL2665
FEATURES             Location/Qualifiers
     source          1..5290
                     /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(1158..1175)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1328..1414
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1506..1533
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1552..1643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1644..1712,1713..2504)
                     /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(1862..1881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2546..3001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2633..2652)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2843..2864
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3112..3699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3600..3619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3853..3870
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4036..4058
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4196..4215
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4395..5273)
                     /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 ttcatcgcta atggcggcga actcaaaccg caggaaaatc
     1021 cgaacgcaca agcgattcag gattaaggta ccaaattcca gaaaagagac gctgaaaagc
     1081 gtcttttttc gttttggtcc tggtctgagg aatggttgta agcttggctg ttttggcgga
     1141 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     1201 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     1261 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     1321 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     1381 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     1441 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     1501 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     1561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1621 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1741 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1861 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     1921 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2041 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2161 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     2221 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2341 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2521 actcatatat actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2581 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2641 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2701 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2761 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2821 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2881 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2941 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct cagcagagcg
     3241 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3361 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3421 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3481 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3541 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3601 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3661 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3721 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3781 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3841 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3901 ctccttacgc atctgtgcgg tatttcacac cgcatagggt catggctgcg ccccgacacc
     3961 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4021 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4081 gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     4141 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     4201 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     4261 gtccggcgta gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt
     4321 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     4381 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     4441 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     4501 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     4561 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     4621 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     4681 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     4741 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     4801 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     4861 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     4921 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     4981 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     5041 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     5101 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     5161 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     5221 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     5281 atactcccgc
//
