LOCUS       sequence-342021-        5312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 2 ring (rings17) LetB mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5312)
  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 5312)
  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: pBEL2005
FEATURES             Location/Qualifiers
     source          1..5312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     438..457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     691..708
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    723..862
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(788..807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     948..970
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(964..1155)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1664..1683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1664..1682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1714..1736
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3933..3951)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3947..3994
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4039..4057)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4125..4229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4230..4298,4299..5090)
                     /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(4448..4467)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5261..5312,1..537)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
       61 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
      121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      601 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      661 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      721 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
      781 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      841 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      901 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      961 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     1021 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     1081 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     1141 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     1201 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     1261 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     1321 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     1381 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     1441 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     1501 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     1561 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     1621 tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata
     1681 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
     1741 catatggctc ttaacacacc acaaattacg ccgacaaaaa agataacagt gagggcaatc
     1801 ggcgaggaac tgccgcgtgg tgattaccaa cgttgcccgc aatgtgacat gctgtttagc
     1861 ctgcccgaga taaattctca tcaaagtgcc tattgtccgc gctgtcaggc aaaaattcgt
     1921 gacgggcgcg actggtcgct aacgcgcctg gcggcaatgg ccttcaccat gctgttgctt
     1981 atgccgtttg cctggggcga accgctgttg catatctggc tgttaggcat tcgtattgac
     2041 gccaacgtta tgcaaggcat ctggcaaatg accaaacagg gcgatgcgat aacggggtcg
     2101 atggtctttt tctgcgttat tggtgccccg ctcattctgg tgacctccat agcttattta
     2161 tggtttggta accgactggg aatgaattta cgtccggtac tgctgatgct tgagcgactt
     2221 aaagagtggg taatgctcga tatctacctg gtcggcattg gcgttgcttc tataaaggta
     2281 caggattatg cccatatcca ggctggtgtc ggcttgttct cttttgtggc gttggtgatt
     2341 ttaacgacgg tgacgttgtc acatcttaat gtcgaagaac tgtgggagcg attttatccg
     2401 cagcgccccg ctacgcgtag ggacgagaaa cttcgtgtct gcctagggtg ccattttacc
     2461 ggctatccag atcagcgtgg tcgctgcccg cgttgccata tcccgctacg cctgcgtcgc
     2521 cgtcatagtc tgcaaaaatg ctgggcggcg ctgttagcgt caatcgtttt attgttacct
     2581 gccaacctgt tgcctatttc tatcatttat ctgaacggag gacggcagga agatacaatt
     2641 ctttccggaa ttatgtcgct ggcaagtagc aacattgcgg ttgcaggaat cgtgtttatc
     2701 gccagtattc tggtcccgtt tactaaagtg atcgtcatgt tcactttact gttgagcatt
     2761 cattttaaat gccagcaagg tttacgcaca cgcattctgt tactgcggat ggtgacctgg
     2821 attggtcgct ggtcgatgct cgacctgttt gtcatatctt taaccatgtc gctgattaat
     2881 cgcgatcaga tcctcgcttt tactatggga ccggctgcgt tttatttcgg cgcagcggta
     2941 attttgacta ttcttgctgt ggaatggctg gacagccgct tactttggga tgcacatgag
     3001 tcaggaaacg cccgcttcga cgactgaagc gcagattaaa aataaacgcc gtatctcacc
     3061 tttctggctg ctgccgttca tcgcgctaat gattgccagt tggctgattt gggacagtta
     3121 tcaggaccgg ggtaataccg tcaccatcga ctttatgtcg gcggatggta ttgttccagg
     3181 ccgtacgcct gttcgttatc agggcgttga agtcggaaca gtgcaggaca tcagcctcag
     3241 cgacgatctt cgtaagattg aagtcaaggt cagcatcaag tccgatatga aagatgcgct
     3301 gcgcgaagag actcagttct ggctggtgac gccaaaagca tcgttggcag gtgtctccgg
     3361 gctggacgcc ctcgtcggtg ggaactatat cggcatgatg ccgggtaaag gtaaagagca
     3421 ggatcacttt gtcgcactcg atacccaacc gaaatatcgg ctggacaatg gcgatttaag
     3481 cattattgtt gaagcgccgg aagccggttc gttaggcatc ggtacgcctg tgctgttccg
     3541 tggtctggaa gtcggtacgg ttacaggaat gacgctgggg acattgtcag atcgcgtgat
     3601 gattgcgatg cgcatcagta aacgctatca acacctggtg cgtaacaatt ccgtcttctg
     3661 gttggcatcg ggttacagtc tggactttgg tctgacgggc ggcgtagtga aaaccggcac
     3721 ctttaaccag tttatccgtg gcggcatcgc cttcgccacg cctccgggta cgccactggc
     3781 accgaaagcc caggaaggca aacacttcct gttgcaggaa agtgaaccga aagagtggcg
     3841 tgaatgggga actgcgcttc ccaaataatg cccacctcga gcagatccgg ctgctaacaa
     3901 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3961 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     4021 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4081 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4141 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4201 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4261 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4321 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4381 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4441 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4501 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4561 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4621 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4681 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4741 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     4801 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4861 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4921 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4981 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5041 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5101 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5161 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5221 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5281 gcgtaatctg ctgcttgcaa acaaaaaaac ca
//
