LOCUS       sequence-309719-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB PLL2 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  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 7040)
  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: pBEL1887
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     176..195
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        176..194
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             226..248
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4173..4191)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4187..4234
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4279..4297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4365..4469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4470..4538,4539..5330)
                     /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(4688..4707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5501..6089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5990..6009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6243..6260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6275..6414
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6340..6359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6500..6522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6516..6707)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
       61 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
      121 catgcgcacc cgtggccagg acccaacgct gcccgagatc tcgatcccgc gaaattaata
      181 cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa
      241 gaaggagata tacatatggc tcttaacaca ccacaaatta cgccgacaaa aaagataaca
      301 gtgagggcaa tcggcgagga actgccgcgt ggtgattacc aacgttgccc gcaatgtgac
      361 atgctgttta gcctgcccga gataaattct catcaaagtg cctattgtcc gcgctgtcag
      421 gcaaaaattc gtgacgggcg cgactggtcg ctaacgcgcc tggcggcaat ggccttcacc
      481 atgctgttgc ttatgccgtt tgcctggggc gaaccgctgt tgcatatctg gctgttaggc
      541 attcgtattg acgccaacgt tatgcaaggc atctggcaaa tgaccaaaca gggcgatgcg
      601 ataacggggt cgatggtctt tttctgcgtt attggtgccc cgctcattct ggtgacctcc
      661 atagcttatt tatggtttgg taaccgactg ggaatgaatt tacgtccggt actgctgatg
      721 cttgagcgac ttaaagagtg ggtaatgctc gatatctacc tggtcggcat tggcgttgct
      781 tctataaagg tacaggatta tgcccatatc caggctggtg tcggcttgtt ctcttttgtg
      841 gcgttggtga ttttaacgac ggtgacgttg tcacatctta atgtcgaaga actgtgggag
      901 cgattttatc cgcagcgccc cgctacgcgt agggacgaga aacttcgtgt ctgcctaggg
      961 tgccatttta ccggctatcc agatcagcgt ggtcgctgcc cgcgttgcca tatcccgcta
     1021 cgcctgcgtc gccgtcatag tctgcaaaaa tgctgggcgg cgctgttagc gtcaatcgtt
     1081 ttattgttac ctgccaacct gttgcctatt tctatcattt atctgaacgg aggacggcag
     1141 gaagatacaa ttctttccgg aattatgtcg ctggcaagta gcaacattgc ggttgcagga
     1201 atcgtgttta tcgccagtat tctggtcccg tttactaaag tgatcgtcat gttcacttta
     1261 ctgttgagca ttcattttaa atgccagcaa ggtttacgca cacgcattct gttactgcgg
     1321 atggtgacct ggattggtcg ctggtcgatg ctcgacctgt ttgtcatatc tttaaccatg
     1381 tcgctgatta atcgcgatca gatcctcgct tttactatgg gaccggctgc gttttatttc
     1441 ggcgcagcgg taattttgac tattcttgct gtggaatggc tggacagccg cttactttgg
     1501 gatgcacatg agtcaggaaa cgcccgcttc gacgactgaa gcgcagatta aaaataaacg
     1561 ccgtatctca cctttctggc tgctgccgtt catcgcgcta atgattgcca gttggctgat
     1621 ttgggacagt tatcaggacc ggggtaatac cgtcaccatc gactttatgt cggcggatgg
     1681 tattgttcca ggccgtacgc ctgttcgtta tcagggcgtt gaagtcggaa cagtgcagga
     1741 catcagcctc agcgacgatc ttcgtaagat tgaagtcaag gtcagcatca agtccgatat
     1801 gaaagatgcg ctgcgcgaag agactcagtt ctggctggtg acgccaaaag catcgttggc
     1861 aggtgtctcc gggctggacg ccctcgtcgg tgggaactat atcggcatga tgcccgggaa
     1921 aggtaaagag caggatcact ttgtcgcact cgatacccaa ccgaaatatc ggctggacaa
     1981 tggcgatctg atgatccacc tgcaagcccc cgatctcggt tcgctgaaca gcggttcatt
     2041 ggtctatttc cgcaagatcc cggtgggaaa agtctacgac tatgccatca atcccaacaa
     2101 gcaaggcgtg gtgattgatg tcctgatcga gcggcgtttt accgatctgg tgaaaaaagg
     2161 tagccgtttc tggaacgttt ccggcgttgg tggggaaagc ttggcggcac tggttaacgg
     2221 tgcgattgcc ttcgattcac cagaagagtc gaaacctgcc gaggcggaag atacctttgg
     2281 tctgtatgaa gatctggccc acagccagcg tggcgtaata ataaaactgg aactgccgag
     2341 tggggccgga ttaaccgccg actcgacgcc gttaatgtat caggggctgg aagtcggaca
     2401 gctgactaaa ctggatttaa atcctggtgg taaagtcaca ggggaaatga ccgttgatcc
     2461 cagcgtcgtt accctgcttc gtgaaaatac ccgcatcgaa ttacgcaacc cgaaattatc
     2521 ccttagcgat gctaatctca gcgccctgct gaccggcaaa accttcgagc tggtgcccgg
     2581 cgatggcgag ccacgcaaag aattcgttgt tgtgccaggc gaaaaagcac tgctgcatga
     2641 acctgatgtt ctgacgctga ccctgaccgc gccggaaagt tacggtattg atgcgggtca
     2701 gccgctcatt cttcacggcg tgcaggtagg ccaggttatc gatcgtaaac tcaccagcaa
     2761 aggcgtcacc tttaccgtcg ccatcgagcc tcagcatcgc gaactggtaa aaggcgatag
     2821 caaatttgtc gtcaacagcc gtgtcgatgt gaaggtgggg ctggatggcg ttgagtttct
     2881 cggtgccagc gcctcagaat ggatcaatgg cgggatccgt attctgccgg gcgataaagg
     2941 tgagatgaaa gccagctatc cactgtatgc caatttggaa aaagcgctgg agaacagcct
     3001 tagcgattta cccaccacaa ccgtgagttt gagtgcagag acgctgccgg atgtgcaggc
     3061 aggatcggta gtgctgtacc gtaaatttga agttggtgaa gtgattaccg tgcgtccgcg
     3121 agctaacgcg tttgacatcg atctgcatat taagccggag tatcgcaacc ttctgaccag
     3181 caatagcgtg ttctgggcag aaggcggggc gaaagttcag ctgaatggta gtggtctgac
     3241 cgtacaggca tccccgctct ccagagctct aaagggagcc attagcttcg acaacctcag
     3301 cggtgccagc gccagtcagc gtaaaggcga taagcgtatt ctgtatgctt ccgaaacagc
     3361 ggcccgtgcg gttggcgggc agattacgct tcacgctttc gatgccggaa aactggcggt
     3421 cgggatgcca attcgctatc tcggtattga catcgggcaa atccagacgc tggatctgat
     3481 taccgcacgc aatgaagtac aggcaaaggc ggtactttac ccggaatatg tccagacctt
     3541 tgctcgcggt ggtacgcgct tctcagtggt cacaccgcaa atttcggcag ctggcgttga
     3601 gcatcttgat actatcctcc agccgtatat caacgtcgaa ccaggccggg gcaatcctcg
     3661 ccgcgacttt gaattacaag aggccaccat tactgattcg cggtacctgg atggcttaag
     3721 cattattgtt gaagcgccgg aagccggttc gttaggcatc ggtacgcctg tgctgttccg
     3781 tggtctggaa gtcggtacgg ttacaggaat gacgctgggg acattgtcag atcgcgtgat
     3841 gattgcgatg cgcatcagta aacgctatca acacctggtg cgtaacaatt ccgtcttctg
     3901 gttggcatcg ggttacagtc tggactttgg tctgacgggc ggcgtagtga aaaccggcac
     3961 ctttaaccag tttatccgtg gcggcatcgc cttcgccacg cctccgggta cgccactggc
     4021 accgaaagcc caggaaggca aacacttcct gttgcaggaa agtgaaccga aagagtggcg
     4081 tgaatgggga actgcgcttc ccaaataatg cccacctcga gcagatccgg ctgctaacaa
     4141 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     4201 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     4261 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4321 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4681 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4981 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     5041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5641 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6241 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     6301 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     6361 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     6421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6541 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     6601 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     6661 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     6721 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     6781 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     6841 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     6901 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     6961 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     7021 accattcatg ttgttgctca
//
