LOCUS       sequence-310592-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB mutant V247N.
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: pBEL1859
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     649..668
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        649..667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             699..721
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4679..4697)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4693..4740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4785..4803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4871..4975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4976..5044,5045..5836)
                     /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(5194..5213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6007..6595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6496..6515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6749..6766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6781..6920
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6846..6865)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7006..7028
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(7022..7073,1..140))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
       61 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
      121 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
      181 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
      241 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
      301 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
      361 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
      421 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
      481 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
      541 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
      601 acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaatta atacgactca
      661 ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
      721 atatacatat ggctcttaac acaccacaaa ttacgccgac aaaaaagata acagtgaggg
      781 caatcggcga ggaactgccg cgtggtgatt accaacgttg cccgcaatgt gacatgctgt
      841 ttagcctgcc cgagataaat tctcatcaaa gtgcctattg tccgcgctgt caggcaaaaa
      901 ttcgtgacgg gcgcgactgg tcgctaacgc gcctggcggc aatggccttc accatgctgt
      961 tgcttatgcc gtttgcctgg ggcgaaccgc tgttgcatat ctggctgtta ggcattcgta
     1021 ttgacgccaa cgttatgcaa ggcatctggc aaatgaccaa acagggcgat gcgataacgg
     1081 ggtcgatggt ctttttctgc gttattggtg ccccgctcat tctggtgacc tccatagctt
     1141 atttatggtt tggtaaccga ctgggaatga atttacgtcc ggtactgctg atgcttgagc
     1201 gacttaaaga gtgggtaatg ctcgatatct acctggtcgg cattggcgtt gcttctataa
     1261 aggtacagga ttatgcccat atccaggctg gtgtcggctt gttctctttt gtggcgttgg
     1321 tgattttaac gacggtgacg ttgtcacatc ttaatgtcga agaactgtgg gagcgatttt
     1381 atccgcagcg ccccgctacg cgtagggacg agaaacttcg tgtctgccta gggtgccatt
     1441 ttaccggcta tccagatcag cgtggtcgct gcccgcgttg ccatatcccg ctacgcctgc
     1501 gtcgccgtca tagtctgcaa aaatgctggg cggcgctgtt agcgtcaatc gttttattgt
     1561 tacctgccaa cctgttgcct atttctatca tttatctgaa cggaggacgg caggaagata
     1621 caattctttc cggaattatg tcgctggcaa gtagcaacat tgcggttgca ggaatcgtgt
     1681 ttatcgccag tattctggtc ccgtttacta aagtgatcgt catgttcact ttactgttga
     1741 gcattcattt taaatgccag caaggtttac gcacacgcat tctgttactg cggatggtga
     1801 cctggattgg tcgctggtcg atgctcgacc tgtttgtcat atctttaacc atgtcgctga
     1861 ttaatcgcga tcagatcctc gcttttacta tgggaccggc tgcgttttat ttcggcgcag
     1921 cggtaatttt gactattctt gctgtggaat ggctggacag ccgcttactt tgggatgcac
     1981 atgagtcagg aaacgcccgc ttcgacgact gaagcgcaga ttaaaaataa acgccgtatc
     2041 tcacctttct ggctgctgcc gttcatcgcg ctaatgattg ccagttggct gatttgggac
     2101 agttatcagg accggggtaa taccgtcacc atcgacttta tgtcggcgga tggtattgtt
     2161 ccaggccgta cgcctgttcg ttatcagggc gttgaagtcg gaacagtgca ggacatcagc
     2221 ctcagcgacg atcttcgtaa gattgaagtc aaggtcagca tcaagtccga tatgaaagat
     2281 gcgctgcgcg aagagactca gttctggctg gtgacgccaa aagcatcgtt ggcaggtgtc
     2341 tccgggctgg acgccctcgt cggtgggaac tatatcggca tgatgccggg taaaggtaaa
     2401 gagcaggatc actttgtcgc actcgatacc caaccgaaat atcggctgga caatggcgat
     2461 ctgatgatcc acctgcaagc ccccgatctc ggttcgctga acagcggttc attggtctat
     2521 ttccgcaaga tcccggtggg aaaagtctac gactatgcca tcaatcccaa caagcaaggc
     2581 gtggtgattg atgtcctgat cgagcggcgt tttaccgatc tggtgaaaaa aggtagccgt
     2641 ttctggaacg tttccggcgt tgatgccaac gtcagtatca gtggcgcgaa ggtgaaactg
     2701 gaaagcctgg cggcactgaa taacggtgcg attgccttcg attcaccaga agagtcgaaa
     2761 cctgccgagg cggaagatac ctttggtctg tatgaagatc tggcccacag ccagcgtggc
     2821 gtaataataa aactggaact gccgagtggg gccggattaa ccgccgactc gacgccgtta
     2881 atgtatcagg ggctggaagt cggacagctg actaaactgg atttaaatcc tggtggtaaa
     2941 gtcacagggg aaatgaccgt tgatcccagc gtcgttaccc tgcttcgtga aaatacccgc
     3001 atcgaattac gcaacccgaa attatccctt agcgatgcta atctcagcgc cctgctgacc
     3061 ggcaaaacct tcgagctggt gcccggcgat ggcgagccac gcaaagagtt cgttgttgtg
     3121 ccaggcgaaa aagcactgct gcatgaacct gatgttctga cgctgaccct gaccgcgccg
     3181 gaaagttacg gtattgatgc gggtcagccg ctcattcttc acggcgtgca ggtaggccag
     3241 gttatcgatc gtaaactcac cagcaaaggc gtcaccttta ccgtcgccat cgagcctcag
     3301 catcgcgaac tggtaaaagg cgatagcaaa tttgtcgtca acagccgtgt cgatgtgaag
     3361 gtggggctgg atggcgttga gtttctcggt gccagcgcct cagaatggat caatggcggg
     3421 atacgtattc tgccgggcga taaaggtgag atgaaagcca gctatccact gtatgccaat
     3481 ttggaaaaag cgctggagaa cagccttagc gatttaccca ccacaaccgt gagtttgagt
     3541 gcagagacgc tgccggatgt gcaggcagga tcggtagtgc tgtaccgtaa atttgaagtt
     3601 ggtgaagtga ttaccgtgcg tccgcgagct aacgcgtttg acatcgatct gcatattaag
     3661 ccggagtatc gcaaccttct gaccagcaat agcgtgttct gggcagaagg cggggcgaaa
     3721 gttcagctga atggtagtgg tctgaccgta caggcatccc cgctctccag agctctaaag
     3781 ggagccatta gcttcgacaa cctcagcggt gccagcgcca gtcagcgtaa aggcgataag
     3841 cgtattctgt atgcttccga aacagcggcc cgtgcggttg gcgggcagat tacgcttcac
     3901 gctttcgatg ccggaaaact ggcggtcggg atgccaattc gctatctcgg tattgacatc
     3961 gggcaaatcc agacgctgga tctgattacc gcacgcaatg aagtacaggc aaaggcggta
     4021 ctttacccgg aatatgtcca gacctttgct cgcggtggta cgcgcttctc agtggtcaca
     4081 ccgcaaattt cggcagctgg cgttgagcat cttgatacta tcctccagcc gtatatcaac
     4141 gtcgaaccag gccggggcaa tcctcgccgc gactttgaat tacaagaggc caccattact
     4201 gattcgcggt acctggatgg cttaagcatt attgttgaag cgccggaagc cggttcgtta
     4261 ggcatcggta cgcctgtgct gttccgtggt ctggaagtcg gtacggttac aggaatgacg
     4321 ctggggacat tgtcagatcg cgtgatgatt gcgatgcgca tcagtaaacg ctatcaacac
     4381 ctggtgcgta acaattccgt cttctggttg gcatcgggtt acagtctgga ctttggtctg
     4441 acgggcggcg tagtgaaaac cggcaccttt aaccagttta tccgtggcgg catcgccttc
     4501 gccacgcctc cgggtacgcc actggcaccg aaagcccagg aaggcaaaca cttcctgttg
     4561 caggaaagtg aaccgaaaga gtggcgtgaa tggggaactg cgcttcccaa ataatgccca
     4621 cctcgagcag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     4681 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     4741 ctgaaaggag gaactatatc cggataattc ttgaagacga aagggcctcg tgatacgcct
     4801 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     5221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     5521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6121 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6781 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     6841 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     6901 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6961 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     7021 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agc
//
