LOCUS       sequence-309760-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB mutant L358N.
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: pBEL1861
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     245..264
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        245..263
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             295..317
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4275..4293)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4289..4336
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4381..4399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4467..4571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4572..4640,4641..5432)
                     /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(4790..4809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5603..6191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6092..6111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6345..6362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6377..6516
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6442..6461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6602..6624
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6618..6809)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
       61 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
      121 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
      181 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct cgatcccgcg
      241 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
      301 taactttaag aaggagatat acatatggct cttaacacac cacaaattac gccgacaaaa
      361 aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca acgttgcccg
      421 caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc ctattgtccg
      481 cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct ggcggcaatg
      541 gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt gcatatctgg
      601 ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat gaccaaacag
      661 ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc gctcattctg
      721 gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt acgtccggta
      781 ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct ggtcggcatt
      841 ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt cggcttgttc
      901 tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa tgtcgaagaa
      961 ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa acttcgtgtc
     1021 tgcctagggt gccattttac cggctatcca gatcagcgtg gtcgctgccc gcgttgccat
     1081 atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc gctgttagcg
     1141 tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta tctgaacgga
     1201 ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag caacattgcg
     1261 gttgcaggaa tcgtgtttat cgccagtatt ctggtcccgt ttactaaagt gatcgtcatg
     1321 ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac acgcattctg
     1381 ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt tgtcatatct
     1441 ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg accggctgcg
     1501 ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct ggacagccgc
     1561 ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag cgcagattaa
     1621 aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa tgattgccag
     1681 ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg actttatgtc
     1741 ggcggatggt attgttccag gccgtacgcc tgttcgttat cagggcgttg aagtcggaac
     1801 agtgcaggac atcagcctca gcgacgatct tcgtaagatt gaagtcaagg tcagcatcaa
     1861 gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga cgccaaaagc
     1921 atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata tcggcatgat
     1981 gccgggtaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac cgaaatatcg
     2041 gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt cgctgaacag
     2101 cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact atgccatcaa
     2161 tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta ccgatctggt
     2221 gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca gtatcagtgg
     2281 cgcgaaggtg aaactggaaa gcctggcggc actggttaac ggtgcgattg ccttcgattc
     2341 accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg aagatctggc
     2401 ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg gattaaccgc
     2461 cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta aactggattt
     2521 aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg ttaccctgct
     2581 tcgtgaaaat acccgcatcg aattacgcaa cccgaaatta tcccttagcg atgctaatct
     2641 cagcgccaac ctgaccggca aaaccttcga gctggtgccc ggcgatggcg agccacgcaa
     2701 agagttcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg ttctgacgct
     2761 gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca ttcttcacgg
     2821 cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca cctttaccgt
     2881 cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg tcgtcaacag
     2941 ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca gcgcctcaga
     3001 atggatcaat ggcgggatac gtattctgcc gggcgataaa ggtgagatga aagccagcta
     3061 tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt tacccaccac
     3121 aaccgtgagt ttgagtgcag agacgctgcc ggatgtgcag gcaggatcgg tagtgctgta
     3181 ccgtaaattt gaagttggtg aagtgattac cgtgcgtccg cgagctaacg cgtttgacat
     3241 cgatctgcat attaagccgg agtatcgcaa ccttctgacc agcaatagcg tgttctgggc
     3301 agaaggcggg gcgaaagttc agctgaatgg tagtggtctg accgtacagg catccccgct
     3361 ctccagagct ctaaagggag ccattagctt cgacaacctc agcggtgcca gcgccagtca
     3421 gcgtaaaggc gataagcgta ttctgtatgc ttccgaaaca gcggcccgtg cggttggcgg
     3481 gcagattacg cttcacgctt tcgatgccgg aaaactggcg gtcgggatgc caattcgcta
     3541 tctcggtatt gacatcgggc aaatccagac gctggatctg attaccgcac gcaatgaagt
     3601 acaggcaaag gcggtacttt acccggaata tgtccagacc tttgctcgcg gtggtacgcg
     3661 cttctcagtg gtcacaccgc aaatttcggc agctggcgtt gagcatcttg atactatcct
     3721 ccagccgtat atcaacgtcg aaccaggccg gggcaatcct cgccgcgact ttgaattaca
     3781 agaggccacc attactgatt cgcggtacct ggatggctta agcattattg ttgaagcgcc
     3841 ggaagccggt tcgttaggca tcggtacgcc tgtgctgttc cgtggtctgg aagtcggtac
     3901 ggttacagga atgacgctgg ggacattgtc agatcgcgtg atgattgcga tgcgcatcag
     3961 taaacgctat caacacctgg tgcgtaacaa ttccgtcttc tggttggcat cgggttacag
     4021 tctggacttt ggtctgacgg gcggcgtagt gaaaaccggc acctttaacc agtttatccg
     4081 tggcggcatc gccttcgcca cgcctccggg tacgccactg gcaccgaaag cccaggaagg
     4141 caaacacttc ctgttgcagg aaagtgaacc gaaagagtgg cgtgaatggg gaactgcgct
     4201 tcccaaataa tgcccacctc gagcagatcc ggctgctaac aaagcccgaa aggaagctga
     4261 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     4321 cttgaggggt tttttgctga aaggaggaac tatatccgga taattcttga agacgaaagg
     4381 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4441 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4561 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4801 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5101 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5281 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5401 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6301 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6361 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6421 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     6481 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     6541 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6601 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     6661 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     6721 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     6781 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     6841 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     6901 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     6961 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     7021 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gca
//
