LOCUS       sequence-320144-        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaE IF1 1-15 aa 
            deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  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 7686)
  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: pBEL2093
FEATURES             Location/Qualifiers
     source          1..7686
                     /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"
     CDS             1893..1910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1923..1943
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3554..3571)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3724..3810
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3902..3929
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3948..4039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4040..4108,4109..4900)
                     /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(4258..4277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4942..5397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5029..5048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5239..5260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5508..6095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5996..6015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6249..6266
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6432..6454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6592..6611
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6791..7669)
                     /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 tagccgaagg gtgaattatg gagcagtctg tggcgaattt agtcgatatg
      361 cgcgatgtca gttttacgcg tggcaatcgc tgcatcttcg ataatatttc cctgaccgtg
      421 ccgcgaggga agatcacggc gatcatgggg ccatcgggca tcggtaaaac gacgctactc
      481 cgtctgattg gcgggcaaat cgcaccagat catggtgaga tccttttcga tggtgagaat
      541 attccggcga tgtctcgttc gcgcctgtat acagtgcgca aacggatgag catgttattt
      601 cagtccgggg cgttgttcac tgatatgaac gtatttgaca acgtcgccta tccactgcgc
      661 gaacataccc aacttcccgc gccattgttg catagtacgg tgatgatgaa gctggaggcc
      721 gtggggctgc gtggagcggc taaactaatg ccttctgaac tttccggtgg gatggcgcgg
      781 cgtgcagcgc tggcgcgtgc gattgcgctg gagccggatc tcatcatgtt tgatgaacct
      841 tttgttgggc aagatcccat caccatgggc gtactggtga agctgatttc tgagctgaac
      901 agcgcgctgg gcgtgacttg tgtggtggtt tctcacgatg tgccggaagt gttaagtatt
      961 gcggatcacg cctggatcct ggcggacaaa aaaattgtcg ctcatggcag tgcccaggcg
     1021 ttgcaggcga atcctgatcc gcgcgtacgt cagtttctgg acgggatagc tgacgggcct
     1081 gttccgttcc gctatcctgc cggcgattat cacgctgatc ttttaccagg gagttaagcc
     1141 actcatgacc ctgagaacgt tcgggcgggc cgggttaatg ttattcaatg cgctggtcgg
     1201 caaaccggaa tttcgcaaac atgcgccgct gctggtgcgc cagctctata atgtcggcgt
     1261 cctgtcgatg ctgattattg tggtttctgg cgtgttcatc ggaatggtgt tggggctgca
     1321 aggttatctg gttctgacca cttatagtgc ggaaaccagt ctgggtatgc tggtggcgtt
     1381 atcgctactg cgtgaactgg ggccggtggt tgccgcgttg ttgtttgccg ggcgtgctgg
     1441 ttcggcgcta accgcagaaa tcggcctgat gcgcgctaca gagcaactct ccagtatgga
     1501 gatgatggcg gtggatccgc tgcgtcgggt tatttctccc cgtttctggg ctggggttat
     1561 ttcattacca ctgttgacgg ttattttcgt cgccgtgggg atctggggcg gatctctggt
     1621 cggcgtcagt tggaaaggca ttgatagcgg gttcttctgg tcggcaatgc aaaatgccgt
     1681 cgactggcgt atggatctgg tcaactgtct gattaagagc gtggtgttcg ccatcacggt
     1741 gacgtggatt tcgttgttta acggctacga cgccatcccg acgtctgccg ggattagccg
     1801 ggcaaccact cgcaccgttg tccactcgtc tctggctgtt ctggggctgg attttgtgct
     1861 gaccgcattg atgtttggga attgagttca tgcatcacca ccatcaccat cagcatcaac
     1921 acgaaaatct gtattttcag gggatgcaaa cgaaaaaaaa tgaaatttgg gtgggtatct
     1981 ttttattagc agcactgctg gcggcgctgt ttgtttgcct gaaggcggcg aacgtgacgt
     2041 ccatacgtac tgaaccgacc tacacgcttt atgcgacgtt cgataacatt ggcggcctga
     2101 aagcgcgctc tccggtcagt attggtggcg ttgttgtggg tcgggtggcg gatattacgc
     2161 tggacccgaa aacctatctg ccgcgcgtaa cgctggaaat tgaacaacgt tataaccaca
     2221 ttccagatac cagttcgctg agcattcgta cttccggcct gctgggggaa caatatctgg
     2281 cattaaacgt cggttttgaa gacccggaac tggggactgc tatcctgaag gatggcgata
     2341 caattcagga cactaagtct gcgatggtgc tggaagatct cattggtcag ttcctttacg
     2401 gtagtaaagg cgatgacaat aagaatagtg gcgatgcgcc agctgctgcg ccaggtaata
     2461 atgaaaccac tgaacctgtg ggtacaacga aataatttca ggagaaccga cgcatgttta
     2521 aacgtttaat gatggtcgct ttgctggtga ttgcacctct gagtgcggca accgcggcag
     2581 accagaccaa tccgtataag ctgatggacg aggcggcgca gaaaacgttc gatcgcctga
     2641 agaatgagca accgcaaatt cgggccaacc cggattatct gcgtaccatt gttgatcagg
     2701 aactgctgcc atacgtacag gtgaaatacg ccggtgcgct ggtgctgggc cagtattaca
     2761 agagtgcgac ccctgctcaa cgtgaagcct actttgccgc tttccgtgag tacctgaagc
     2821 aggcttacgg tcaggcgctg gcgatgtatc acggtcaaac ctatcagatt gcgccagaac
     2881 agccgctggg cgataaaacc attgtgccta ttcgcgttac cattattgac ccgaatggcc
     2941 gtccgccggt gcgtctggac ttccagtggc gtaaaaactc ccagacgggc aattggcagg
     3001 cttacgacat gattgctgaa ggcgtcagta tgatcaccac caaacaaaac gagtggggaa
     3061 cgctgctgcg taccaaaggt atcgacggcc tgactgcgca actgaaatcg atttctcaac
     3121 agaaaatcac tctggaagag aaaaaataat gagcgagtca ctgagctgga tgcagacggg
     3181 tgacacgctg gcgttatccg gagagctgga tcaggacgtt ttgctaccgc tttgggaaat
     3241 gcgtgaggaa gcggtgaagg ggattacctg catcgatctt agccgtgtct cccgcgtgga
     3301 tacgggggga ctggcactgc tgctccatct tatcgatctg gcgaaaaagc agggcaacaa
     3361 tgtgacgctt cagggggtaa acgacaaagt gtataccctg gcgaaattgt ataatttgcc
     3421 tgctgatgtt ctgcctcgtt aaggtaccaa attccagaaa agagacgctg aaaagcgtct
     3481 tttttcgttt tggtcctggt ctgaggaatg gttgtaagct tggctgtttt ggcggatgag
     3541 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     3601 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     3661 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     3721 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3781 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     3841 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3901 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4021 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4081 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4141 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4201 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4261 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4321 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4381 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4441 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4501 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4561 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4621 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4681 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4741 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4801 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4861 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4921 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4981 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5041 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5101 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     5161 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5221 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5281 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5341 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     5401 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggctcagc agagcgcaga
     5641 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5701 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5761 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5821 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5881 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5941 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6001 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6061 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6121 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6181 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6241 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6301 ttacgcatct gtgcggtatt tcacaccgca tagggtcatg gctgcgcccc gacacccgcc
     6361 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6421 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6481 gaggcagaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6541 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6601 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6661 ggcgtagagg atctgctcat gtttgacagc ttatcatcga tgcataatgt gcctgtcaaa
     6721 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     6781 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     6841 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     6901 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     6961 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     7021 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     7081 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     7141 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     7201 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     7261 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     7321 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     7381 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     7441 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     7501 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     7561 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     7621 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     7681 tcccgc
//
