LOCUS       sequence-385686-        7545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of MlaF-MlaE-MlaDdelta141-183-MlaC-MlaB .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7545)
  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 7545)
  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: pBEL2766
FEATURES             Location/Qualifiers
     source          1..7545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..299
                     /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     222..241
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3418..3435)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3588..3674
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3766..3793
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3812..3903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3904..3972,3973..4764)
                     /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(4122..4141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4806..5261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4893..4912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5103..5124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5372..5959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5860..5879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6113..6130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6296..6318
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6456..6475
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6655..7533)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
       61 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
      121 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
      181 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
      241 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
      301 cccgtttttt tgggctagcc gaagggtgaa ttatggagca gtctgtggcg aatttagtcg
      361 atatgcgcga tgtcagtttt acgcgtggca atcgctgcat cttcgataat atttccctga
      421 ccgtgccgcg agggaagatc acggcgatca tggggccatc gggcatcggt aaaacgacgc
      481 tactccgtct gattggcggg caaatcgcac cagatcatgg tgagatcctt ttcgatggtg
      541 agaatattcc ggcgatgtct cgttcgcgcc tgtatacagt gcgcaaacgg atgagcatgt
      601 tatttcagtc cggggcgttg ttcactgata tgaacgtatt tgacaacgtc gcctatccac
      661 tgcgcgaaca tacccaactt cccgcgccat tgttgcatag tacggtgatg atgaagctgg
      721 aggccgtggg gctgcgtgga gcggctaaac taatgccttc tgaactttcc ggtgggatgg
      781 cgcggcgtgc agcgctggcg cgtgcgattg cgctggagcc ggatctcatc atgtttgatg
      841 aaccttttgt tgggcaagat cccatcacca tgggcgtact ggtgaagctg atttctgagc
      901 tgaacagcgc gctgggcgtg acttgtgtgg tggtttctca cgatgtgccg gaagtgttaa
      961 gtattgcgga tcacgcctgg atcctggcgg acaaaaaaat tgtcgctcat ggcagtgccc
     1021 aggcgttgca ggcgaatcct gatccgcgcg tacgtcagtt tctggacggg atagctgacg
     1081 ggcctgttcc gttccgctat cctgccggcg attatcacgc tgatctttta ccagggagtt
     1141 aagccactca tgctgttaaa tgcgctggcg tcgctcggac ataaagggat taaaaccctg
     1201 agaacgttcg ggcgggccgg gttaatgtta ttcaatgcgc tggtcggcaa accggaattt
     1261 cgcaaacatg cgccgctgct ggtgcgccag ctctataatg tcggcgtcct gtcgatgctg
     1321 attattgtgg tttctggcgt gttcatcgga atggtgttgg ggctgcaagg ttatctggtt
     1381 ctgaccactt atagtgcgga aaccagtctg ggtatgctgg tggcgttatc gctactgcgt
     1441 gaactggggc cggtggttgc cgcgttgttg tttgccgggc gtgctggttc ggcgctaacc
     1501 gcagaaatcg gcctgatgcg cgctacagag caactctcca gtatggagat gatggcggtg
     1561 gatccgctgc gtcgggttat ttctccccgt ttctgggctg gggttatttc attaccactg
     1621 ttgacggtta ttttcgtcgc cgtggggatc tggggcggat ctctggtcgg cgtcagttgg
     1681 aaaggcattg atagcgggtt cttctggtcg gcaatgcaaa atgccgtcga ctggcgtatg
     1741 gatctggtca actgtctgat taagagcgtg gtgttcgcca tcacggtgac gtggatttcg
     1801 ttgtttaacg gctacgacgc catcccgacg tctgccggga ttagccgggc aaccactcgc
     1861 accgttgtcc actcgtctct ggctgttctg gggctggatt ttgtgctgac cgcattgatg
     1921 tttgggaatt gagttcatgc aaacgaaaaa aaatgaaatt tgggtgggta tctttttatt
     1981 agcagcactg ctggcggcgc tgtttgtttg cctgaaggcg gcgaacgtga cgtccatacg
     2041 tactgaaccg acctacacgc tttatgcgac gttcgataac attggcggcc tgaaagcgcg
     2101 ctctccggtc agtattggtg gcgttgttgt gggtcgggtg gcggatatta cgctggaccc
     2161 gaaaacctat ctgccgcgcg taacgctgga aattgaacaa cgttataacc acattccaga
     2221 taccagttcg ctgagcattc gtacttccgg cctgctgggg gaacaatatc tggcattaaa
     2281 cgtcggtttt gaagacccgg aactggggac tgctatcctg aaggatggcg atacaattca
     2341 ggacactaag tctgcgtaat ttcaggagaa ccgacgcatg tttaaacgtt taatgatggt
     2401 cgctttgctg gtgattgcac ctctgagtgc ggcaaccgcg gcagaccaga ccaatccgta
     2461 taagctgatg gacgaggcgg cgcagaaaac gttcgatcgc ctgaagaatg agcaaccgca
     2521 aattcgggcc aacccggatt atctgcgtac cattgttgat caggaactgc tgccatacgt
     2581 acaggtgaaa tacgccggtg cgctggtgct gggccagtat tacaagagtg cgacccctgc
     2641 tcaacgtgaa gcctactttg ccgctttccg tgagtacctg aagcaggctt acggtcaggc
     2701 gctggcgatg tatcacggtc aaacctatca gattgcgcca gaacagccgc tgggcgataa
     2761 aaccattgtg cctattcgcg ttaccattat tgacccgaat ggccgtccgc cggtgcgtct
     2821 ggacttccag tggcgtaaaa actcccagac gggcaattgg caggcttacg acatgattgc
     2881 tgaaggcgtc agtatgatca ccaccaaaca aaacgagtgg ggaacgctgc tgcgtaccaa
     2941 aggtatcgac ggcctgactg cgcaactgaa atcgatttct caacagaaaa tcactctgga
     3001 agagaaaaaa taatgagcga gtcactgagc tggatgcaga cgggtgacac gctggcgtta
     3061 tccggagagc tggatcagga cgttttgcta ccgctttggg aaatgcgtga ggaagcggtg
     3121 aaggggatta cctgcatcga tcttagccgt gtctcccgcg tggatacggg gggactggca
     3181 ctgctgctcc atcttatcga tctggcgaaa aagcagggca acaatgtgac gcttcagggg
     3241 gtaaacgaca aagtgtatac cctggcgaaa ttgtataatt tgcctgctga tgttctgcct
     3301 cgttaaggta ccaaattcca gaaaagagac gctgaaaagc gtcttttttc gttttggtcc
     3361 tggtctgagg aatggttgta agcttggctg ttttggcgga tgagagaaga ttttcagcct
     3421 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     3481 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     3541 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     3601 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     3661 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     3721 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     3781 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     3841 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3901 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3961 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4021 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4081 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4141 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4201 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4261 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4321 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4381 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4441 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4501 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4561 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4621 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4681 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4741 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4801 gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4861 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4921 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4981 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5041 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5101 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     5161 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5221 atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga
     5281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5461 taccaactct ttttccgaag gtaactggct cagcagagcg cagataccaa atactgtcct
     5521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6121 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6181 tatttcacac cgcatagggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     6241 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     6301 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gaaggagatg
     6361 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     6421 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     6481 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc
     6541 tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat
     6601 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga
     6661 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     6721 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     6781 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     6841 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga
     6901 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     6961 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     7021 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     7081 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     7141 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     7201 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     7261 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     7321 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     7381 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     7441 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     7501 cggcagcagg ggatcatttt gcgcttcagc catacttttc atact
//
