LOCUS       sequence-326370-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for MlaD (A20F).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7728)
  AUTHORS   Coudray N, Isom GL, MacRae MR, Saiduddin MN, Bhabha G, Ekiert DC
  TITLE     Structure of bacterial phospholipid transporter MlaFEDB with 
            substrate bound.
  JOURNAL   Elife. 2020 Nov 25;9. pii: 62518. doi: 10.7554/eLife.62518.
  PUBMED    33236984
REFERENCE   2  (bases 1 to 7728)
  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: "Elife."; 
            date: "2020-11-25"; volume: "9. pii: 62518. doi"; pages: " 
            10.7554/eLife.62518"
COMMENT     Sequence Label: pBEL2296
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(151..168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(322..909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(402..421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(1020..1475)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1157..1178)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1369..1388
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(1517..2308,2309..2377))
                     /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     2140..2159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2378..2469)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2488..2515)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2607..2693)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2846..2863
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(5261..5281)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5294..5311)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(6165..6449)
                     /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     complement(6223..6242)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6476..7354
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(7534..7553)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7691..7713)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
       61 ggtgtcgggg cgcagccatg accctatgcg gtgtgaaata ccgcacagat gcgtaaggag
      121 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      721 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
      781 cgctctgctg agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1021 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1081 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
     1141 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     1201 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     1261 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1321 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     1381 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1441 acccgccgcg cttaatgcgc cgctacaggg cgcgtaaatc aatctaaagt atatatgagt
     1501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1801 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     1861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2101 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     2161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2461 ataaacaaaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     2521 atttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     2581 gcaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     2641 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
     2701 gcagttccct actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac
     2761 ttctgagttc ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg
     2821 ttttatcaga ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca
     2881 tccgccaaaa cagccaagct tacaaccatt cctcagacca ggaccaaaac gaaaaaagac
     2941 gcttttcagc gtctcttttc tggaatttgg taccttaacg aggcagaaca tcagcaggca
     3001 aattatacaa tttcgccagg gtatacactt tgtcgtttac cccctgaagc gtcacattgt
     3061 tgccctgctt tttcgccaga tcgataagat ggagcagcag tgccagtccc cccgtatcca
     3121 cgcgggagac acggctaaga tcgatgcagg taatcccctt caccgcttcc tcacgcattt
     3181 cccaaagcgg tagcaaaacg tcctgatcca gctctccgga taacgccagc gtgtcacccg
     3241 tctgcatcca gctcagtgac tcgctcatta ttttttctct tccagagtga ttttctgttg
     3301 agaaatcgat ttcagttgcg cagtcaggcc gtcgatacct ttggtacgca gcagcgttcc
     3361 ccactcgttt tgtttggtgg tgatcatact gacgccttca gcaatcatgt cgtaagcctg
     3421 ccaattgccc gtctgggagt ttttacgcca ctggaagtcc agacgcaccg gcggacggcc
     3481 attcgggtca ataatggtaa cgcgaatagg cacaatggtt ttatcgccca gcggctgttc
     3541 tggcgcaatc tgataggttt gaccgtgata catcgccagc gcctgaccgt aagcctgctt
     3601 caggtactca cggaaagcgg caaagtaggc ttcacgttga gcaggggtcg cactcttgta
     3661 atactggccc agcaccagcg caccggcgta tttcacctgt acgtatggca gcagttcctg
     3721 atcaacaatg gtacgcagat aatccgggtt ggcccgaatt tgcggttgct cattcttcag
     3781 gcgatcgaac gttttctgcg ccgcctcgtc catcagctta tacggattgg tctggtctgc
     3841 cgcggttgcc gcactcagag gtgcaatcac cagcaaagcg accatcatta aacgtttaaa
     3901 catgcgtcgg ttctcctgaa attatttcgt tgtacccaca ggttcagtgg tttcattatt
     3961 acctggcgca gcagctggcg catcgccact attcttattg tcatcgcctt tactaccgta
     4021 aaggaactga ccaatgagat cttccagcac catcgcagac ttagtgtcct gaattgtatc
     4081 gccatccttc aggatagcag tccccagttc cgggtcttca aaaccgacgt ttaatgccag
     4141 atattgttcc cccagcaggc cggaagtacg aatgctcagc gaactggtat ctggaatgtg
     4201 gttataacgt tgttcaattt ccagcgttac gcgcggcaga taggttttcg ggtccagcgt
     4261 aatatccgcc acccgaccca caacaacgcc accaatactg accggagagc gcgctttcag
     4321 gccgccaatg ttatcgaacg tcgcataaag cgtgtaggtc ggttcagtac gtatggacgt
     4381 cacgttcgcc gccttcaggc aaacaaacag cgcaaacagc agtgctgcta ataaaaagat
     4441 acccacccaa atttcatttt ttttcgtttg catgaactca attcccaaac atcaatgcgg
     4501 tcagcacaaa atccagcccc agaacagcca gagacgagtg gacaacggtg cgagtggttg
     4561 cccggctaat cccggcagac gtcgggatgg cgtcgtagcc gttaaacaac gaaatccacg
     4621 tcaccgtgat ggcgaacacc acgctcttaa tcagacagtt gaccagatcc atacgccagt
     4681 cgacggcatt ttgcattgcc gaccagaaga acccgctatc aatgcctttc caactgacgc
     4741 cgaccagaga tccgccccag atccccacgg cgacgaaaat aaccgtcaac agtggtaatg
     4801 aaataacccc agcccagaaa cggggagaaa taacccgacg cagcggatcc accgccatca
     4861 tctccatact ggagagttgc tctgtagcgc gcatcaggcc gatttctgcg gttagcgccg
     4921 aaccagcacg cccggcaaac aacaacgcgg caaccaccgg ccccagttca cgcagtagcg
     4981 ataacgccac cagcataccc agactggttt ccgcactata agtggtcaga accagataac
     5041 cttgcagccc caacaccatt ccgatgaaca cgccagaaac cacaataatc agcatcgaca
     5101 ggacgccgac attatagagc tggcgcacca gcagcggcgc atgtttgcga aattccggtt
     5161 tgccgaccag cgcattgaat aacattaacc cggcccgccc gaacgttctc agggttttaa
     5221 tccctttatg tccgagcgac gccagcgcat ttaacagcat cccctgaaaa tacagatttt
     5281 cgtgttgatg ctgatggtga tggtggtgat gcatgagtgg cttaactccc tggtaaaaga
     5341 tcagcgtgat aatcgccggc aggatagcgg aacggaacag gcccgtcagc tatcccgtcc
     5401 agaaactgac gtacgcgcgg atcaggattc gcctgcaacg cctgggcact gccatgagcg
     5461 acaatttttt tgtccgccag gatccaggcg tgatccgcaa tacttaacac ttccggcaca
     5521 tcgtgagaaa ccaccacaca agtcacgccc agcgcgctgt tcagctcaga aatcagcttc
     5581 accagtacgc ccatggtgat gggatcttgc ccaacaaaag gttcatcaaa catgatgaga
     5641 tccggctcca gcgcaatcgc acgcgccagc gctgcacgcc gcgccatccc accggaaagt
     5701 tcagaaggca ttagtttagc cgctccacgc agccccacgg cctccagctt catcatcacc
     5761 gtactatgca acaatggcgc gggaagttgg gtatgttcgc gcagtggata ggcgacgttg
     5821 tcaaatacgt tcatatcagt gaacaacgcc ccggactgaa ataacatgct catccgtttg
     5881 cgcactgtat acaggcgcga acgagacatc gccggaatat tctcaccatc gaaaaggatc
     5941 tcaccatgat ctggtgcgat ttgcccgcca atcagacgga gtagcgtcgt tttaccgatg
     6001 cccgatggcc ccatgatcgc cgtgatcttc cctcgcggca cggtcaggga aatattatcg
     6061 aagatgcagc gattgccacg cgtaaaactg acatcgcgca tatcgactaa attcgccaca
     6121 gactgctcca taattcaccc ttcggctagc ccaaaaaaac gggtatggag aaacagtaga
     6181 gagttgcgat aaaaagcgtc aggtaggatc cgctaatctt atggataaaa atgctatggc
     6241 atagcaaagt gtgacgccgt gcaaataatc aatgtggact tttctgccgt gattatagac
     6301 acttttgtta cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa tgcttttaat
     6361 aagcggggtt accggtttgg ttagcgagaa gagccagtaa aagacgcagt gacggcaatg
     6421 tctgatgcaa tatggacaat tggtttcttc tctgaatggc gggagtatga aaagtatggc
     6481 tgaagcgcaa aatgatcccc tgctgccggg atactcgttt aatgcccatc tggtggcggg
     6541 tttaacgccg attgaggcca acggttatct cgattttttt atcgaccgac cgctgggaat
     6601 gaaaggttat attctcaatc tcaccattcg cggtcagggg gtggtgaaaa atcagggacg
     6661 agaatttgtt tgccgaccgg gtgatatttt gctgttcccg ccaggagaga ttcatcacta
     6721 cggtcgtcat ccggaggctc gcgaatggta tcaccagtgg gtttactttc gtccgcgcgc
     6781 ctactggcat gaatggctta actggccgtc aatatttgcc aatacggggt tctttcgccc
     6841 ggatgaagcg caccagccgc atttcagcga cctgtttggg caaatcatta acgccgggca
     6901 aggggaaggg cgctattcgg agctgctggc gataaatctg cttgagcaat tgttactgcg
     6961 gcgcatggaa gcgattaacg agtcgctcca tccaccgatg gataatcggg tacgcgaggc
     7021 ttgtcagtac atcagcgatc acctggcaga cagcaatttt gatatcgcca gcgtcgcaca
     7081 gcatgtttgc ttgtcgccgt cgcgtctgtc acatcttttc cgccagcagt tagggattag
     7141 cgtcttaagc tggcgcgagg accaacgtat cagccaggcg aagctgcttt tgagcaccac
     7201 ccggatgcct atcgccaccg tcggtcgcaa tgttggtttt gacgatcaac tctatttctc
     7261 gcgggtattt aaaaaatgca ccggggccag cccgagcgag ttccgtgccg gttgtgaaga
     7321 aaaagtgaat gatgtagccg tcaagttgtc ataattggta acgaatcaga caattgacgg
     7381 cttgacggag tagcataggg tttgcagaat ccctgcttcg tccatttgac aggcacatta
     7441 tgcatcgatg ataagctgtc aaacatgagc agatcctcta cgccggacgc atcgtggccg
     7501 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     7561 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     7621 gccccgtggc cgggggactg ttgggcgcca tctccttctg cctcgcgcgt ttcggtgatg
     7681 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagctt
//
