LOCUS       sequence-383737-        5272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaA(delta238-251).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5272)
  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 5272)
  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: pBEL2512
FEATURES             Location/Qualifiers
     source          1..5272
                     /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(1140..1157)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1310..1396
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1488..1515
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1534..1625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1626..1694,1695..2486)
                     /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(1844..1863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2528..2983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2615..2634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2825..2846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3094..3681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3582..3601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3835..3852
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4018..4040
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4178..4197
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4377..5255)
                     /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 cgaactaagg taccaaattc cagaaaagag acgctgaaaa gcgtcttttt tcgttttggt
     1081 cctggtctga ggaatggttg taagcttggc tgttttggcg gatgagagaa gattttcagc
     1141 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     1201 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     1261 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     1321 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1381 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     1441 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     1501 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     1561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1621 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1681 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1741 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1801 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1861 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1921 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1981 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2041 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2101 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2161 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2221 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2281 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2341 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2401 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2461 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2521 ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2581 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2641 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2701 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     2761 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2821 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     2881 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2941 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa
     3001 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3181 gctaccaact ctttttccga aggtaactgg ctcagcagag cgcagatacc aaatactgtc
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3841 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcatagg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3961 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4021 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga
     4081 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     4141 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     4201 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     4261 gctcatgttt gacagcttat catcgatgca taatgtgcct gtcaaatgga cgaagcaggg
     4321 attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat
     4381 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
     4441 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
     4501 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
     4561 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
     4621 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
     4681 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
     4741 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     4801 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     4861 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     4921 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     4981 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     5041 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     5101 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     5161 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     5221 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gc
//
