LOCUS       sequence-309752-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB mutant L243N.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   Isom GL, Coudray N, MacRae MR, McManus CT, Ekiert DC, Bhabha G
  TITLE     LetB Structure Reveals a Tunnel for Lipid Transport across the 
            Bacterial Envelope.
  JOURNAL   Cell. 2020 Apr 30;181(3):653-664.e19. doi: 
            10.1016/j.cell.2020.03.030.
  PUBMED    32359438
REFERENCE   2  (bases 1 to 7073)
  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"; doi: 
            "10.1016/j.cell.2020.03"; journalName: "Cell"; date: "2020-04-30- 
            30"; volume: "181"; issue: "3"; pages: "653-664.e19"
COMMENT     Sequence Label: pBEL1857
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     270..289
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        270..288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             320..342
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4300..4318)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4314..4361
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4406..4424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4492..4596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4597..4665,4666..5457)
                     /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(4815..4834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5628..6216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6117..6136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6370..6387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6402..6541
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6467..6486)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6627..6649
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6643..6834)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
       61 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
      121 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
      181 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
      241 cgctgcccga gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
      301 gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata tggctcttaa
      361 cacaccacaa attacgccga caaaaaagat aacagtgagg gcaatcggcg aggaactgcc
      421 gcgtggtgat taccaacgtt gcccgcaatg tgacatgctg tttagcctgc ccgagataaa
      481 ttctcatcaa agtgcctatt gtccgcgctg tcaggcaaaa attcgtgacg ggcgcgactg
      541 gtcgctaacg cgcctggcgg caatggcctt caccatgctg ttgcttatgc cgtttgcctg
      601 gggcgaaccg ctgttgcata tctggctgtt aggcattcgt attgacgcca acgttatgca
      661 aggcatctgg caaatgacca aacagggcga tgcgataacg gggtcgatgg tctttttctg
      721 cgttattggt gccccgctca ttctggtgac ctccatagct tatttatggt ttggtaaccg
      781 actgggaatg aatttacgtc cggtactgct gatgcttgag cgacttaaag agtgggtaat
      841 gctcgatatc tacctggtcg gcattggcgt tgcttctata aaggtacagg attatgccca
      901 tatccaggct ggtgtcggct tgttctcttt tgtggcgttg gtgattttaa cgacggtgac
      961 gttgtcacat cttaatgtcg aagaactgtg ggagcgattt tatccgcagc gccccgctac
     1021 gcgtagggac gagaaacttc gtgtctgcct agggtgccat tttaccggct atccagatca
     1081 gcgtggtcgc tgcccgcgtt gccatatccc gctacgcctg cgtcgccgtc atagtctgca
     1141 aaaatgctgg gcggcgctgt tagcgtcaat cgttttattg ttacctgcca acctgttgcc
     1201 tatttctatc atttatctga acggaggacg gcaggaagat acaattcttt ccggaattat
     1261 gtcgctggca agtagcaaca ttgcggttgc aggaatcgtg tttatcgcca gtattctggt
     1321 cccgtttact aaagtgatcg tcatgttcac tttactgttg agcattcatt ttaaatgcca
     1381 gcaaggttta cgcacacgca ttctgttact gcggatggtg acctggattg gtcgctggtc
     1441 gatgctcgac ctgtttgtca tatctttaac catgtcgctg attaatcgcg atcagatcct
     1501 cgcttttact atgggaccgg ctgcgtttta tttcggcgca gcggtaattt tgactattct
     1561 tgctgtggaa tggctggaca gccgcttact ttgggatgca catgagtcag gaaacgcccg
     1621 cttcgacgac tgaagcgcag attaaaaata aacgccgtat ctcacctttc tggctgctgc
     1681 cgttcatcgc gctaatgatt gccagttggc tgatttggga cagttatcag gaccggggta
     1741 ataccgtcac catcgacttt atgtcggcgg atggtattgt tccaggccgt acgcctgttc
     1801 gttatcaggg cgttgaagtc ggaacagtgc aggacatcag cctcagcgac gatcttcgta
     1861 agattgaagt caaggtcagc atcaagtccg atatgaaaga tgcgctgcgc gaagagactc
     1921 agttctggct ggtgacgcca aaagcatcgt tggcaggtgt ctccgggctg gacgccctcg
     1981 tcggtgggaa ctatatcggc atgatgccgg gtaaaggtaa agagcaggat cactttgtcg
     2041 cactcgatac ccaaccgaaa tatcggctgg acaatggcga tctgatgatc cacctgcaag
     2101 cccccgatct cggttcgctg aacagcggtt cattggtcta tttccgcaag atcccggtgg
     2161 gaaaagtcta cgactatgcc atcaatccca acaagcaagg cgtggtgatt gatgtcctga
     2221 tcgagcggcg ttttaccgat ctggtgaaaa aaggtagccg tttctggaac gtttccggcg
     2281 ttgatgccaa cgtcagtatc agtggcgcga aggtgaaact ggaaagcaat gcggcactgg
     2341 ttaacggtgc gattgccttc gattcaccag aagagtcgaa acctgccgag gcggaagata
     2401 cctttggtct gtatgaagat ctggcccaca gccagcgtgg cgtaataata aaactggaac
     2461 tgccgagtgg ggccggatta accgccgact cgacgccgtt aatgtatcag gggctggaag
     2521 tcggacagct gactaaactg gatttaaatc ctggtggtaa agtcacaggg gaaatgaccg
     2581 ttgatcccag cgtcgttacc ctgcttcgtg aaaatacccg catcgaatta cgcaacccga
     2641 aattatccct tagcgatgct aatctcagcg ccctgctgac cggcaaaacc ttcgagctgg
     2701 tgcccggcga tggcgagcca cgcaaagagt tcgttgttgt gccaggcgaa aaagcactgc
     2761 tgcatgaacc tgatgttctg acgctgaccc tgaccgcgcc ggaaagttac ggtattgatg
     2821 cgggtcagcc gctcattctt cacggcgtgc aggtaggcca ggttatcgat cgtaaactca
     2881 ccagcaaagg cgtcaccttt accgtcgcca tcgagcctca gcatcgcgaa ctggtaaaag
     2941 gcgatagcaa atttgtcgtc aacagccgtg tcgatgtgaa ggtggggctg gatggcgttg
     3001 agtttctcgg tgccagcgcc tcagaatgga tcaatggcgg gatacgtatt ctgccgggcg
     3061 ataaaggtga gatgaaagcc agctatccac tgtatgccaa tttggaaaaa gcgctggaga
     3121 acagccttag cgatttaccc accacaaccg tgagtttgag tgcagagacg ctgccggatg
     3181 tgcaggcagg atcggtagtg ctgtaccgta aatttgaagt tggtgaagtg attaccgtgc
     3241 gtccgcgagc taacgcgttt gacatcgatc tgcatattaa gccggagtat cgcaaccttc
     3301 tgaccagcaa tagcgtgttc tgggcagaag gcggggcgaa agttcagctg aatggtagtg
     3361 gtctgaccgt acaggcatcc ccgctctcca gagctctaaa gggagccatt agcttcgaca
     3421 acctcagcgg tgccagcgcc agtcagcgta aaggcgataa gcgtattctg tatgcttccg
     3481 aaacagcggc ccgtgcggtt ggcgggcaga ttacgcttca cgctttcgat gccggaaaac
     3541 tggcggtcgg gatgccaatt cgctatctcg gtattgacat cgggcaaatc cagacgctgg
     3601 atctgattac cgcacgcaat gaagtacagg caaaggcggt actttacccg gaatatgtcc
     3661 agacctttgc tcgcggtggt acgcgcttct cagtggtcac accgcaaatt tcggcagctg
     3721 gcgttgagca tcttgatact atcctccagc cgtatatcaa cgtcgaacca ggccggggca
     3781 atcctcgccg cgactttgaa ttacaagagg ccaccattac tgattcgcgg tacctggatg
     3841 gcttaagcat tattgttgaa gcgccggaag ccggttcgtt aggcatcggt acgcctgtgc
     3901 tgttccgtgg tctggaagtc ggtacggtta caggaatgac gctggggaca ttgtcagatc
     3961 gcgtgatgat tgcgatgcgc atcagtaaac gctatcaaca cctggtgcgt aacaattccg
     4021 tcttctggtt ggcatcgggt tacagtctgg actttggtct gacgggcggc gtagtgaaaa
     4081 ccggcacctt taaccagttt atccgtggcg gcatcgcctt cgccacgcct ccgggtacgc
     4141 cactggcacc gaaagcccag gaaggcaaac acttcctgtt gcaggaaagt gaaccgaaag
     4201 agtggcgtga atggggaact gcgcttccca aataatgccc acctcgagca gatccggctg
     4261 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     4321 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     4381 ccggataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4441 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4501 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4561 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4621 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4681 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4741 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4801 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4861 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4921 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4981 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5041 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5101 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5161 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5221 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5281 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5341 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5401 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5461 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5761 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6241 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6301 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6361 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     6421 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     6481 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     6541 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6601 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6661 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     6721 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     6781 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     6841 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     6901 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     6961 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     7021 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cga
//
