LOCUS       sequence-309757-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB mutant L355N.
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: pBEL1860
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             262..284
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4242..4260)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4256..4303
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4348..4366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4434..4538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4539..4607,4608..5399)
                     /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(4757..4776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5570..6158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6059..6078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6312..6329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6344..6483
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6409..6428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6569..6591
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6585..6776)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
       61 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
      121 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
      181 aacgctgccc gagatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
      241 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatggctctt
      301 aacacaccac aaattacgcc gacaaaaaag ataacagtga gggcaatcgg cgaggaactg
      361 ccgcgtggtg attaccaacg ttgcccgcaa tgtgacatgc tgtttagcct gcccgagata
      421 aattctcatc aaagtgccta ttgtccgcgc tgtcaggcaa aaattcgtga cgggcgcgac
      481 tggtcgctaa cgcgcctggc ggcaatggcc ttcaccatgc tgttgcttat gccgtttgcc
      541 tggggcgaac cgctgttgca tatctggctg ttaggcattc gtattgacgc caacgttatg
      601 caaggcatct ggcaaatgac caaacagggc gatgcgataa cggggtcgat ggtctttttc
      661 tgcgttattg gtgccccgct cattctggtg acctccatag cttatttatg gtttggtaac
      721 cgactgggaa tgaatttacg tccggtactg ctgatgcttg agcgacttaa agagtgggta
      781 atgctcgata tctacctggt cggcattggc gttgcttcta taaaggtaca ggattatgcc
      841 catatccagg ctggtgtcgg cttgttctct tttgtggcgt tggtgatttt aacgacggtg
      901 acgttgtcac atcttaatgt cgaagaactg tgggagcgat tttatccgca gcgccccgct
      961 acgcgtaggg acgagaaact tcgtgtctgc ctagggtgcc attttaccgg ctatccagat
     1021 cagcgtggtc gctgcccgcg ttgccatatc ccgctacgcc tgcgtcgccg tcatagtctg
     1081 caaaaatgct gggcggcgct gttagcgtca atcgttttat tgttacctgc caacctgttg
     1141 cctatttcta tcatttatct gaacggagga cggcaggaag atacaattct ttccggaatt
     1201 atgtcgctgg caagtagcaa cattgcggtt gcaggaatcg tgtttatcgc cagtattctg
     1261 gtcccgttta ctaaagtgat cgtcatgttc actttactgt tgagcattca ttttaaatgc
     1321 cagcaaggtt tacgcacacg cattctgtta ctgcggatgg tgacctggat tggtcgctgg
     1381 tcgatgctcg acctgtttgt catatcttta accatgtcgc tgattaatcg cgatcagatc
     1441 ctcgctttta ctatgggacc ggctgcgttt tatttcggcg cagcggtaat tttgactatt
     1501 cttgctgtgg aatggctgga cagccgctta ctttgggatg cacatgagtc aggaaacgcc
     1561 cgcttcgacg actgaagcgc agattaaaaa taaacgccgt atctcacctt tctggctgct
     1621 gccgttcatc gcgctaatga ttgccagttg gctgatttgg gacagttatc aggaccgggg
     1681 taataccgtc accatcgact ttatgtcggc ggatggtatt gttccaggcc gtacgcctgt
     1741 tcgttatcag ggcgttgaag tcggaacagt gcaggacatc agcctcagcg acgatcttcg
     1801 taagattgaa gtcaaggtca gcatcaagtc cgatatgaaa gatgcgctgc gcgaagagac
     1861 tcagttctgg ctggtgacgc caaaagcatc gttggcaggt gtctccgggc tggacgccct
     1921 cgtcggtggg aactatatcg gcatgatgcc gggtaaaggt aaagagcagg atcactttgt
     1981 cgcactcgat acccaaccga aatatcggct ggacaatggc gatctgatga tccacctgca
     2041 agcccccgat ctcggttcgc tgaacagcgg ttcattggtc tatttccgca agatcccggt
     2101 gggaaaagtc tacgactatg ccatcaatcc caacaagcaa ggcgtggtga ttgatgtcct
     2161 gatcgagcgg cgttttaccg atctggtgaa aaaaggtagc cgtttctgga acgtttccgg
     2221 cgttgatgcc aacgtcagta tcagtggcgc gaaggtgaaa ctggaaagcc tggcggcact
     2281 ggttaacggt gcgattgcct tcgattcacc agaagagtcg aaacctgccg aggcggaaga
     2341 tacctttggt ctgtatgaag atctggccca cagccagcgt ggcgtaataa taaaactgga
     2401 actgccgagt ggggccggat taaccgccga ctcgacgccg ttaatgtatc aggggctgga
     2461 agtcggacag ctgactaaac tggatttaaa tcctggtggt aaagtcacag gggaaatgac
     2521 cgttgatccc agcgtcgtta ccctgcttcg tgaaaatacc cgcatcgaat tacgcaaccc
     2581 gaaattatcc cttagcgatg ctaataatag cgccctgctg accggcaaaa ccttcgagct
     2641 ggtgcccggc gatggcgagc cacgcaaaga gttcgttgtt gtgccaggcg aaaaagcact
     2701 gctgcatgaa cctgatgttc tgacgctgac cctgaccgcg ccggaaagtt acggtattga
     2761 tgcgggtcag ccgctcattc ttcacggcgt gcaggtaggc caggttatcg atcgtaaact
     2821 caccagcaaa ggcgtcacct ttaccgtcgc catcgagcct cagcatcgcg aactggtaaa
     2881 aggcgatagc aaatttgtcg tcaacagccg tgtcgatgtg aaggtggggc tggatggcgt
     2941 tgagtttctc ggtgccagcg cctcagaatg gatcaatggc gggatacgta ttctgccggg
     3001 cgataaaggt gagatgaaag ccagctatcc actgtatgcc aatttggaaa aagcgctgga
     3061 gaacagcctt agcgatttac ccaccacaac cgtgagtttg agtgcagaga cgctgccgga
     3121 tgtgcaggca ggatcggtag tgctgtaccg taaatttgaa gttggtgaag tgattaccgt
     3181 gcgtccgcga gctaacgcgt ttgacatcga tctgcatatt aagccggagt atcgcaacct
     3241 tctgaccagc aatagcgtgt tctgggcaga aggcggggcg aaagttcagc tgaatggtag
     3301 tggtctgacc gtacaggcat ccccgctctc cagagctcta aagggagcca ttagcttcga
     3361 caacctcagc ggtgccagcg ccagtcagcg taaaggcgat aagcgtattc tgtatgcttc
     3421 cgaaacagcg gcccgtgcgg ttggcgggca gattacgctt cacgctttcg atgccggaaa
     3481 actggcggtc gggatgccaa ttcgctatct cggtattgac atcgggcaaa tccagacgct
     3541 ggatctgatt accgcacgca atgaagtaca ggcaaaggcg gtactttacc cggaatatgt
     3601 ccagaccttt gctcgcggtg gtacgcgctt ctcagtggtc acaccgcaaa tttcggcagc
     3661 tggcgttgag catcttgata ctatcctcca gccgtatatc aacgtcgaac caggccgggg
     3721 caatcctcgc cgcgactttg aattacaaga ggccaccatt actgattcgc ggtacctgga
     3781 tggcttaagc attattgttg aagcgccgga agccggttcg ttaggcatcg gtacgcctgt
     3841 gctgttccgt ggtctggaag tcggtacggt tacaggaatg acgctgggga cattgtcaga
     3901 tcgcgtgatg attgcgatgc gcatcagtaa acgctatcaa cacctggtgc gtaacaattc
     3961 cgtcttctgg ttggcatcgg gttacagtct ggactttggt ctgacgggcg gcgtagtgaa
     4021 aaccggcacc tttaaccagt ttatccgtgg cggcatcgcc ttcgccacgc ctccgggtac
     4081 gccactggca ccgaaagccc aggaaggcaa acacttcctg ttgcaggaaa gtgaaccgaa
     4141 agagtggcgt gaatggggaa ctgcgcttcc caaataatgc ccacctcgag cagatccggc
     4201 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     4261 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     4321 atccggataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4381 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     4801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     5101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6181 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6241 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6301 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     6361 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     6421 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     6481 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6541 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6601 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     6661 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     6721 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     6781 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     6841 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     6901 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     6961 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     7021 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cga
//
