LOCUS       sequence-383734-        5239 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Complementation of MlaA(delta227-251).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  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 5239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pBEL2513
FEATURES             Location/Qualifiers
     source          1..5239
                     /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(1107..1124)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1277..1363
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1455..1482
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1501..1592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1593..1661,1662..2453)
                     /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(1811..1830)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2495..2950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2582..2601)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2792..2813
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3061..3648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3549..3568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3802..3819
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3985..4007
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4145..4164
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4344..5222)
                     /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 attaaggtac caaattccag aaaagagacg
     1021 ctgaaaagcg tcttttttcg ttttggtcct ggtctgagga atggttgtaa gcttggctgt
     1081 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1141 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1201 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1261 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1321 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1381 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1441 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1501 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1561 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1621 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1681 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1741 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1801 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1861 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1921 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1981 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2041 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2101 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     2161 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2221 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2281 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2341 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2401 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2461 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     2521 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2581 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2641 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2701 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2761 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2821 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2881 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2941 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3001 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3061 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3121 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctc
     3181 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3241 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3301 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3361 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3421 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3481 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3541 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3601 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3661 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3721 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3781 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3841 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggtc atggctgcgc
     3901 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3961 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4021 caccgaaacg cgcgaggcag aaggagatgg cgcccaacag tcccccggcc acggggcctg
     4081 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     4141 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     4201 ccacgatgcg tccggcgtag aggatctgct catgtttgac agcttatcat cgatgcataa
     4261 tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg
     4321 tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct
     4381 tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag
     4441 aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg
     4501 gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta
     4561 atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt
     4621 gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc
     4681 tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc
     4741 agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc
     4801 ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg
     4861 cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg
     4921 cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag
     4981 tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt
     5041 ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt
     5101 cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc
     5161 gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc
     5221 atacttttca tactcccgc
//
