LOCUS       sequence-320147-        7656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaE IF1 1-25 aa 
            deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7656)
  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 7656)
  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: pBEL2132
FEATURES             Location/Qualifiers
     source          1..7656
                     /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             1863..1880
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1893..1913
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3524..3541)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3694..3780
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3872..3899
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3918..4009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4010..4078,4079..4870)
                     /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(4228..4247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4912..5367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4999..5018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5209..5230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5478..6065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5966..5985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6219..6236
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6402..6424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6562..6581
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6761..7639)
                     /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 actcatgatg ttattcaatg cgctggtcgg caaaccggaa tttcgcaaac atgcgccgct
     1201 gctggtgcgc cagctctata atgtcggcgt cctgtcgatg ctgattattg tggtttctgg
     1261 cgtgttcatc ggaatggtgt tggggctgca aggttatctg gttctgacca cttatagtgc
     1321 ggaaaccagt ctgggtatgc tggtggcgtt atcgctactg cgtgaactgg ggccggtggt
     1381 tgccgcgttg ttgtttgccg ggcgtgctgg ttcggcgcta accgcagaaa tcggcctgat
     1441 gcgcgctaca gagcaactct ccagtatgga gatgatggcg gtggatccgc tgcgtcgggt
     1501 tatttctccc cgtttctggg ctggggttat ttcattacca ctgttgacgg ttattttcgt
     1561 cgccgtgggg atctggggcg gatctctggt cggcgtcagt tggaaaggca ttgatagcgg
     1621 gttcttctgg tcggcaatgc aaaatgccgt cgactggcgt atggatctgg tcaactgtct
     1681 gattaagagc gtggtgttcg ccatcacggt gacgtggatt tcgttgttta acggctacga
     1741 cgccatcccg acgtctgccg ggattagccg ggcaaccact cgcaccgttg tccactcgtc
     1801 tctggctgtt ctggggctgg attttgtgct gaccgcattg atgtttggga attgagttca
     1861 tgcatcacca ccatcaccat cagcatcaac acgaaaatct gtattttcag gggatgcaaa
     1921 cgaaaaaaaa tgaaatttgg gtgggtatct ttttattagc agcactgctg gcggcgctgt
     1981 ttgtttgcct gaaggcggcg aacgtgacgt ccatacgtac tgaaccgacc tacacgcttt
     2041 atgcgacgtt cgataacatt ggcggcctga aagcgcgctc tccggtcagt attggtggcg
     2101 ttgttgtggg tcgggtggcg gatattacgc tggacccgaa aacctatctg ccgcgcgtaa
     2161 cgctggaaat tgaacaacgt tataaccaca ttccagatac cagttcgctg agcattcgta
     2221 cttccggcct gctgggggaa caatatctgg cattaaacgt cggttttgaa gacccggaac
     2281 tggggactgc tatcctgaag gatggcgata caattcagga cactaagtct gcgatggtgc
     2341 tggaagatct cattggtcag ttcctttacg gtagtaaagg cgatgacaat aagaatagtg
     2401 gcgatgcgcc agctgctgcg ccaggtaata atgaaaccac tgaacctgtg ggtacaacga
     2461 aataatttca ggagaaccga cgcatgttta aacgtttaat gatggtcgct ttgctggtga
     2521 ttgcacctct gagtgcggca accgcggcag accagaccaa tccgtataag ctgatggacg
     2581 aggcggcgca gaaaacgttc gatcgcctga agaatgagca accgcaaatt cgggccaacc
     2641 cggattatct gcgtaccatt gttgatcagg aactgctgcc atacgtacag gtgaaatacg
     2701 ccggtgcgct ggtgctgggc cagtattaca agagtgcgac ccctgctcaa cgtgaagcct
     2761 actttgccgc tttccgtgag tacctgaagc aggcttacgg tcaggcgctg gcgatgtatc
     2821 acggtcaaac ctatcagatt gcgccagaac agccgctggg cgataaaacc attgtgccta
     2881 ttcgcgttac cattattgac ccgaatggcc gtccgccggt gcgtctggac ttccagtggc
     2941 gtaaaaactc ccagacgggc aattggcagg cttacgacat gattgctgaa ggcgtcagta
     3001 tgatcaccac caaacaaaac gagtggggaa cgctgctgcg taccaaaggt atcgacggcc
     3061 tgactgcgca actgaaatcg atttctcaac agaaaatcac tctggaagag aaaaaataat
     3121 gagcgagtca ctgagctgga tgcagacggg tgacacgctg gcgttatccg gagagctgga
     3181 tcaggacgtt ttgctaccgc tttgggaaat gcgtgaggaa gcggtgaagg ggattacctg
     3241 catcgatctt agccgtgtct cccgcgtgga tacgggggga ctggcactgc tgctccatct
     3301 tatcgatctg gcgaaaaagc agggcaacaa tgtgacgctt cagggggtaa acgacaaagt
     3361 gtataccctg gcgaaattgt ataatttgcc tgctgatgtt ctgcctcgtt aaggtaccaa
     3421 attccagaaa agagacgctg aaaagcgtct tttttcgttt tggtcctggt ctgaggaatg
     3481 gttgtaagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     3541 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     3601 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     3661 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3721 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3781 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     3841 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     3901 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     4261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     4561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct
     4921 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4981 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5041 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5101 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     5161 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5221 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     5281 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5341 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     5401 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5461 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5521 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5581 ccgaaggtaa ctggctcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6181 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6241 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6301 tagggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6361 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6421 gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc
     6481 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6541 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6601 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc
     6661 ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     6721 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     6781 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     6841 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     6901 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     6961 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     7021 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     7081 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     7141 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     7201 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     7261 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     7321 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     7381 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     7441 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     7501 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     7561 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     7621 cattttgcgc ttcagccata cttttcatac tcccgc
//
