LOCUS       sequence-326631-        7728 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Complementation plasmid for MlaD (I12F).
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 Apr 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: pBEL2291
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(47..133)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     286..303
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2701..2721)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2734..2751)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(3605..3889)
                     /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(3663..3682)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3916..4794
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(4974..4993)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5131..5153)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5319..5336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5490..6077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5570..5589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(6188..6643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6325..6346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6537..6556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(6685..7476,7477..7545))
                     /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     7308..7327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7546..7637)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(7656..7683)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 accctccggg ccgttgcttc gcaacgttca aatccgctcc cggcggattt gtcctactca
       61 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
      121 ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagaccc cacactacca
      181 tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac caccgcgcta
      241 ctgccgccag gcaaattctg ttttatcaga ccgcttctgc gttctgattt aatctgtatc
      301 aggctgaaaa tcttctctca tccgccaaaa cagccaagct tacaaccatt cctcagacca
      361 ggaccaaaac gaaaaaagac gcttttcagc gtctcttttc tggaatttgg taccttaacg
      421 aggcagaaca tcagcaggca aattatacaa tttcgccagg gtatacactt tgtcgtttac
      481 cccctgaagc gtcacattgt tgccctgctt tttcgccaga tcgataagat ggagcagcag
      541 tgccagtccc cccgtatcca cgcgggagac acggctaaga tcgatgcagg taatcccctt
      601 caccgcttcc tcacgcattt cccaaagcgg tagcaaaacg tcctgatcca gctctccgga
      661 taacgccagc gtgtcacccg tctgcatcca gctcagtgac tcgctcatta ttttttctct
      721 tccagagtga ttttctgttg agaaatcgat ttcagttgcg cagtcaggcc gtcgatacct
      781 ttggtacgca gcagcgttcc ccactcgttt tgtttggtgg tgatcatact gacgccttca
      841 gcaatcatgt cgtaagcctg ccaattgccc gtctgggagt ttttacgcca ctggaagtcc
      901 agacgcaccg gcggacggcc attcgggtca ataatggtaa cgcgaatagg cacaatggtt
      961 ttatcgccca gcggctgttc tggcgcaatc tgataggttt gaccgtgata catcgccagc
     1021 gcctgaccgt aagcctgctt caggtactca cggaaagcgg caaagtaggc ttcacgttga
     1081 gcaggggtcg cactcttgta atactggccc agcaccagcg caccggcgta tttcacctgt
     1141 acgtatggca gcagttcctg atcaacaatg gtacgcagat aatccgggtt ggcccgaatt
     1201 tgcggttgct cattcttcag gcgatcgaac gttttctgcg ccgcctcgtc catcagctta
     1261 tacggattgg tctggtctgc cgcggttgcc gcactcagag gtgcaatcac cagcaaagcg
     1321 accatcatta aacgtttaaa catgcgtcgg ttctcctgaa attatttcgt tgtacccaca
     1381 ggttcagtgg tttcattatt acctggcgca gcagctggcg catcgccact attcttattg
     1441 tcatcgcctt tactaccgta aaggaactga ccaatgagat cttccagcac catcgcagac
     1501 ttagtgtcct gaattgtatc gccatccttc aggatagcag tccccagttc cgggtcttca
     1561 aaaccgacgt ttaatgccag atattgttcc cccagcaggc cggaagtacg aatgctcagc
     1621 gaactggtat ctggaatgtg gttataacgt tgttcaattt ccagcgttac gcgcggcaga
     1681 taggttttcg ggtccagcgt aatatccgcc acccgaccca caacaacgcc accaatactg
     1741 accggagagc gcgctttcag gccgccaatg ttatcgaacg tcgcataaag cgtgtaggtc
     1801 ggttcagtac gtatggacgt cacgttcgcc gccttcaggc aaacaaacag cgccgccagc
     1861 agtgctgcta ataaaaacgc acccacccaa atttcatttt ttttcgtttg catgaactca
     1921 attcccaaac atcaatgcgg tcagcacaaa atccagcccc agaacagcca gagacgagtg
     1981 gacaacggtg cgagtggttg cccggctaat cccggcagac gtcgggatgg cgtcgtagcc
     2041 gttaaacaac gaaatccacg tcaccgtgat ggcgaacacc acgctcttaa tcagacagtt
     2101 gaccagatcc atacgccagt cgacggcatt ttgcattgcc gaccagaaga acccgctatc
     2161 aatgcctttc caactgacgc cgaccagaga tccgccccag atccccacgg cgacgaaaat
     2221 aaccgtcaac agtggtaatg aaataacccc agcccagaaa cggggagaaa taacccgacg
     2281 cagcggatcc accgccatca tctccatact ggagagttgc tctgtagcgc gcatcaggcc
     2341 gatttctgcg gttagcgccg aaccagcacg cccggcaaac aacaacgcgg caaccaccgg
     2401 ccccagttca cgcagtagcg ataacgccac cagcataccc agactggttt ccgcactata
     2461 agtggtcaga accagataac cttgcagccc caacaccatt ccgatgaaca cgccagaaac
     2521 cacaataatc agcatcgaca ggacgccgac attatagagc tggcgcacca gcagcggcgc
     2581 atgtttgcga aattccggtt tgccgaccag cgcattgaat aacattaacc cggcccgccc
     2641 gaacgttctc agggttttaa tccctttatg tccgagcgac gccagcgcat ttaacagcat
     2701 cccctgaaaa tacagatttt cgtgttgatg ctgatggtga tggtggtgat gcatgagtgg
     2761 cttaactccc tggtaaaaga tcagcgtgat aatcgccggc aggatagcgg aacggaacag
     2821 gcccgtcagc tatcccgtcc agaaactgac gtacgcgcgg atcaggattc gcctgcaacg
     2881 cctgggcact gccatgagcg acaatttttt tgtccgccag gatccaggcg tgatccgcaa
     2941 tacttaacac ttccggcaca tcgtgagaaa ccaccacaca agtcacgccc agcgcgctgt
     3001 tcagctcaga aatcagcttc accagtacgc ccatggtgat gggatcttgc ccaacaaaag
     3061 gttcatcaaa catgatgaga tccggctcca gcgcaatcgc acgcgccagc gctgcacgcc
     3121 gcgccatccc accggaaagt tcagaaggca ttagtttagc cgctccacgc agccccacgg
     3181 cctccagctt catcatcacc gtactatgca acaatggcgc gggaagttgg gtatgttcgc
     3241 gcagtggata ggcgacgttg tcaaatacgt tcatatcagt gaacaacgcc ccggactgaa
     3301 ataacatgct catccgtttg cgcactgtat acaggcgcga acgagacatc gccggaatat
     3361 tctcaccatc gaaaaggatc tcaccatgat ctggtgcgat ttgcccgcca atcagacgga
     3421 gtagcgtcgt tttaccgatg cccgatggcc ccatgatcgc cgtgatcttc cctcgcggca
     3481 cggtcaggga aatattatcg aagatgcagc gattgccacg cgtaaaactg acatcgcgca
     3541 tatcgactaa attcgccaca gactgctcca taattcaccc ttcggctagc ccaaaaaaac
     3601 gggtatggag aaacagtaga gagttgcgat aaaaagcgtc aggtaggatc cgctaatctt
     3661 atggataaaa atgctatggc atagcaaagt gtgacgccgt gcaaataatc aatgtggact
     3721 tttctgccgt gattatagac acttttgtta cgcgtttttg tcatggcttt ggtcccgctt
     3781 tgttacagaa tgcttttaat aagcggggtt accggtttgg ttagcgagaa gagccagtaa
     3841 aagacgcagt gacggcaatg tctgatgcaa tatggacaat tggtttcttc tctgaatggc
     3901 gggagtatga aaagtatggc tgaagcgcaa aatgatcccc tgctgccggg atactcgttt
     3961 aatgcccatc tggtggcggg tttaacgccg attgaggcca acggttatct cgattttttt
     4021 atcgaccgac cgctgggaat gaaaggttat attctcaatc tcaccattcg cggtcagggg
     4081 gtggtgaaaa atcagggacg agaatttgtt tgccgaccgg gtgatatttt gctgttcccg
     4141 ccaggagaga ttcatcacta cggtcgtcat ccggaggctc gcgaatggta tcaccagtgg
     4201 gtttactttc gtccgcgcgc ctactggcat gaatggctta actggccgtc aatatttgcc
     4261 aatacggggt tctttcgccc ggatgaagcg caccagccgc atttcagcga cctgtttggg
     4321 caaatcatta acgccgggca aggggaaggg cgctattcgg agctgctggc gataaatctg
     4381 cttgagcaat tgttactgcg gcgcatggaa gcgattaacg agtcgctcca tccaccgatg
     4441 gataatcggg tacgcgaggc ttgtcagtac atcagcgatc acctggcaga cagcaatttt
     4501 gatatcgcca gcgtcgcaca gcatgtttgc ttgtcgccgt cgcgtctgtc acatcttttc
     4561 cgccagcagt tagggattag cgtcttaagc tggcgcgagg accaacgtat cagccaggcg
     4621 aagctgcttt tgagcaccac ccggatgcct atcgccaccg tcggtcgcaa tgttggtttt
     4681 gacgatcaac tctatttctc gcgggtattt aaaaaatgca ccggggccag cccgagcgag
     4741 ttccgtgccg gttgtgaaga aaaagtgaat gatgtagccg tcaagttgtc ataattggta
     4801 acgaatcaga caattgacgg cttgacggag tagcataggg tttgcagaat ccctgcttcg
     4861 tccatttgac aggcacatta tgcatcgatg ataagctgtc aaacatgagc agatcctcta
     4921 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     4981 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     5041 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttctg
     5101 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5161 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5221 tgttggcggg tgtcggggcg cagccatgac cctatgcggt gtgaaatacc gcacagatgc
     5281 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5401 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5941 ggtatctgcg ctctgctgag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6001 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6061 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6121 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6181 tccttttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6241 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6301 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6361 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6421 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6481 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6541 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6601 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaatcaa tctaaagtat
     6661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     7021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     7321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7621 ttagaaaaat aaacaaaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     7681 tctgcttaat ttgatgcctg gcagtttatg gcgggcgtcc tgcccgcc
//
