LOCUS       sequence-320155-        7614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaE IF1 1-39 aa 
            deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7614)
  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 7614)
  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: pBEL2133
FEATURES             Location/Qualifiers
     source          1..7614
                     /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             1821..1838
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1851..1871
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3482..3499)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3652..3738
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3830..3857
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3876..3967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3968..4036,4037..4828)
                     /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(4186..4205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4870..5325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4957..4976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5167..5188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5436..6023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5924..5943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6177..6194
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6360..6382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6520..6539
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6719..7597)
                     /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 actcatgcat gcgccgctgc tggtgcgcca gctctataat gtcggcgtcc tgtcgatgct
     1201 gattattgtg gtttctggcg tgttcatcgg aatggtgttg gggctgcaag gttatctggt
     1261 tctgaccact tatagtgcgg aaaccagtct gggtatgctg gtggcgttat cgctactgcg
     1321 tgaactgggg ccggtggttg ccgcgttgtt gtttgccggg cgtgctggtt cggcgctaac
     1381 cgcagaaatc ggcctgatgc gcgctacaga gcaactctcc agtatggaga tgatggcggt
     1441 ggatccgctg cgtcgggtta tttctccccg tttctgggct ggggttattt cattaccact
     1501 gttgacggtt attttcgtcg ccgtggggat ctggggcgga tctctggtcg gcgtcagttg
     1561 gaaaggcatt gatagcgggt tcttctggtc ggcaatgcaa aatgccgtcg actggcgtat
     1621 ggatctggtc aactgtctga ttaagagcgt ggtgttcgcc atcacggtga cgtggatttc
     1681 gttgtttaac ggctacgacg ccatcccgac gtctgccggg attagccggg caaccactcg
     1741 caccgttgtc cactcgtctc tggctgttct ggggctggat tttgtgctga ccgcattgat
     1801 gtttgggaat tgagttcatg catcaccacc atcaccatca gcatcaacac gaaaatctgt
     1861 attttcaggg gatgcaaacg aaaaaaaatg aaatttgggt gggtatcttt ttattagcag
     1921 cactgctggc ggcgctgttt gtttgcctga aggcggcgaa cgtgacgtcc atacgtactg
     1981 aaccgaccta cacgctttat gcgacgttcg ataacattgg cggcctgaaa gcgcgctctc
     2041 cggtcagtat tggtggcgtt gttgtgggtc gggtggcgga tattacgctg gacccgaaaa
     2101 cctatctgcc gcgcgtaacg ctggaaattg aacaacgtta taaccacatt ccagatacca
     2161 gttcgctgag cattcgtact tccggcctgc tgggggaaca atatctggca ttaaacgtcg
     2221 gttttgaaga cccggaactg gggactgcta tcctgaagga tggcgataca attcaggaca
     2281 ctaagtctgc gatggtgctg gaagatctca ttggtcagtt cctttacggt agtaaaggcg
     2341 atgacaataa gaatagtggc gatgcgccag ctgctgcgcc aggtaataat gaaaccactg
     2401 aacctgtggg tacaacgaaa taatttcagg agaaccgacg catgtttaaa cgtttaatga
     2461 tggtcgcttt gctggtgatt gcacctctga gtgcggcaac cgcggcagac cagaccaatc
     2521 cgtataagct gatggacgag gcggcgcaga aaacgttcga tcgcctgaag aatgagcaac
     2581 cgcaaattcg ggccaacccg gattatctgc gtaccattgt tgatcaggaa ctgctgccat
     2641 acgtacaggt gaaatacgcc ggtgcgctgg tgctgggcca gtattacaag agtgcgaccc
     2701 ctgctcaacg tgaagcctac tttgccgctt tccgtgagta cctgaagcag gcttacggtc
     2761 aggcgctggc gatgtatcac ggtcaaacct atcagattgc gccagaacag ccgctgggcg
     2821 ataaaaccat tgtgcctatt cgcgttacca ttattgaccc gaatggccgt ccgccggtgc
     2881 gtctggactt ccagtggcgt aaaaactccc agacgggcaa ttggcaggct tacgacatga
     2941 ttgctgaagg cgtcagtatg atcaccacca aacaaaacga gtggggaacg ctgctgcgta
     3001 ccaaaggtat cgacggcctg actgcgcaac tgaaatcgat ttctcaacag aaaatcactc
     3061 tggaagagaa aaaataatga gcgagtcact gagctggatg cagacgggtg acacgctggc
     3121 gttatccgga gagctggatc aggacgtttt gctaccgctt tgggaaatgc gtgaggaagc
     3181 ggtgaagggg attacctgca tcgatcttag ccgtgtctcc cgcgtggata cggggggact
     3241 ggcactgctg ctccatctta tcgatctggc gaaaaagcag ggcaacaatg tgacgcttca
     3301 gggggtaaac gacaaagtgt ataccctggc gaaattgtat aatttgcctg ctgatgttct
     3361 gcctcgttaa ggtaccaaat tccagaaaag agacgctgaa aagcgtcttt tttcgttttg
     3421 gtcctggtct gaggaatggt tgtaagcttg gctgttttgg cggatgagag aagattttca
     3481 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     3541 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     3601 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     3661 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     3721 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     3781 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     3841 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     3901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3961 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4021 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4081 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4141 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4201 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4261 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4321 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4381 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4441 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4501 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4561 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4621 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4681 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4741 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4801 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4861 gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4921 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4981 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5041 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5101 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5161 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     5221 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5281 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg
     5341 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5401 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5461 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5521 gagctaccaa ctctttttcc gaaggtaact ggctcagcag agcgcagata ccaaatactg
     5581 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5641 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5701 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5761 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5821 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5881 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5941 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6001 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6061 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6121 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6181 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     6241 gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa cacccgctga
     6301 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6361 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga
     6421 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     6481 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     6541 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     6601 ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag
     6661 ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt
     6721 atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg
     6781 ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa
     6841 cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc
     6901 tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat
     6961 gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc
     7021 tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg
     7081 gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta
     7141 tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca
     7201 ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata
     7261 ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt
     7321 gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca
     7381 gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac
     7441 cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat
     7501 cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt
     7561 atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc ccgc
//
