LOCUS       sequence-270781-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB mutant L246N.
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: pBEL1858
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4092..4110)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4106..4153
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4198..4216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4284..4388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4389..4457,4458..5249)
                     /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(4607..4626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5420..6008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5909..5928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6162..6179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6194..6333
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6259..6278)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6419..6441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6435..6626)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatggctctt aacacaccac aaattacgcc gacaaaaaag
      181 ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa
      241 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      301 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      361 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      421 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      481 gatgcgataa cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg
      541 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      601 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      661 gttgcttcta taaaggtaca ggattatgcc catatccagg ctggtgtcgg cttgttctct
      721 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
      781 tgggagcgat tttatccgca gcgccccgct acgcgtaggg acgagaaact tcgtgtctgc
      841 ctagggtgcc attttaccgg ctatccagat cagcgtggtc gctgcccgcg ttgccatatc
      901 ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct gggcggcgct gttagcgtca
      961 atcgttttat tgttacctgc caacctgttg cctatttcta tcatttatct gaacggagga
     1021 cggcaggaag atacaattct ttccggaatt atgtcgctgg caagtagcaa cattgcggtt
     1081 gcaggaatcg tgtttatcgc cagtattctg gtcccgttta ctaaagtgat cgtcatgttc
     1141 actttactgt tgagcattca ttttaaatgc cagcaaggtt tacgcacacg cattctgtta
     1201 ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg acctgtttgt catatcttta
     1261 accatgtcgc tgattaatcg cgatcagatc ctcgctttta ctatgggacc ggctgcgttt
     1321 tatttcggcg cagcggtaat tttgactatt cttgctgtgg aatggctgga cagccgctta
     1381 ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa
     1441 taaacgccgt atctcacctt tctggctgct gccgttcatc gcgctaatga ttgccagttg
     1501 gctgatttgg gacagttatc aggaccgggg taataccgtc accatcgact ttatgtcggc
     1561 ggatggtatt gttccaggcc gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt
     1621 gcaggacatc agcctcagcg acgatcttcg taagattgaa gtcaaggtca gcatcaagtc
     1681 cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg ctggtgacgc caaaagcatc
     1741 gttggcaggt gtctccgggc tggacgccct cgtcggtggg aactatatcg gcatgatgcc
     1801 gggtaaaggt aaagagcagg atcactttgt cgcactcgat acccaaccga aatatcggct
     1861 ggacaatggc gatctgatga tccacctgca agcccccgat ctcggttcgc tgaacagcgg
     1921 ttcattggtc tatttccgca agatcccggt gggaaaagtc tacgactatg ccatcaatcc
     1981 caacaagcaa ggcgtggtga ttgatgtcct gatcgagcgg cgttttaccg atctggtgaa
     2041 aaaaggtagc cgtttctgga acgtttccgg cgttgatgcc aacgtcagta tcagtggcgc
     2101 gaaggtgaaa ctggaaagcc tggcggcaaa cgttaacggt gcgattgcct tcgattcacc
     2161 agaagagtcg aaacctgccg aggcggaaga tacctttggt ctgtatgaag atctggccca
     2221 cagccagcgt ggcgtaataa taaaactgga actgccgagt ggggccggat taaccgccga
     2281 ctcgacgccg ttaatgtatc aggggctgga agtcggacag ctgactaaac tggatttaaa
     2341 tcctggtggt aaagtcacag gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg
     2401 tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc cttagcgatg ctaatctcag
     2461 cgccctgctg accggcaaaa ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga
     2521 gttcgttgtt gtgccaggcg aaaaagcact gctgcatgaa cctgatgttc tgacgctgac
     2581 cctgaccgcg ccggaaagtt acggtattga tgcgggtcag ccgctcattc ttcacggcgt
     2641 gcaggtaggc caggttatcg atcgtaaact caccagcaaa ggcgtcacct ttaccgtcgc
     2701 catcgagcct cagcatcgcg aactggtaaa aggcgatagc aaatttgtcg tcaacagccg
     2761 tgtcgatgtg aaggtggggc tggatggcgt tgagtttctc ggtgccagcg cctcagaatg
     2821 gatcaatggc gggatacgta ttctgccggg cgataaaggt gagatgaaag ccagctatcc
     2881 actgtatgcc aatttggaaa aagcgctgga gaacagcctt agcgatttac ccaccacaac
     2941 cgtgagtttg agtgcagaga cgctgccgga tgtgcaggca ggatcggtag tgctgtaccg
     3001 taaatttgaa gttggtgaag tgattaccgt gcgtccgcga gctaacgcgt ttgacatcga
     3061 tctgcatatt aagccggagt atcgcaacct tctgaccagc aatagcgtgt tctgggcaga
     3121 aggcggggcg aaagttcagc tgaatggtag tggtctgacc gtacaggcat ccccgctctc
     3181 cagagctcta aagggagcca ttagcttcga caacctcagc ggtgccagcg ccagtcagcg
     3241 taaaggcgat aagcgtattc tgtatgcttc cgaaacagcg gcccgtgcgg ttggcgggca
     3301 gattacgctt cacgctttcg atgccggaaa actggcggtc gggatgccaa ttcgctatct
     3361 cggtattgac atcgggcaaa tccagacgct ggatctgatt accgcacgca atgaagtaca
     3421 ggcaaaggcg gtactttacc cggaatatgt ccagaccttt gctcgcggtg gtacgcgctt
     3481 ctcagtggtc acaccgcaaa tttcggcagc tggcgttgag catcttgata ctatcctcca
     3541 gccgtatatc aacgtcgaac caggccgggg caatcctcgc cgcgactttg aattacaaga
     3601 ggccaccatt actgattcgc ggtacctgga tggcttaagc attattgttg aagcgccgga
     3661 agccggttcg ttaggcatcg gtacgcctgt gctgttccgt ggtctggaag tcggtacggt
     3721 tacaggaatg acgctgggga cattgtcaga tcgcgtgatg attgcgatgc gcatcagtaa
     3781 acgctatcaa cacctggtgc gtaacaattc cgtcttctgg ttggcatcgg gttacagtct
     3841 ggactttggt ctgacgggcg gcgtagtgaa aaccggcacc tttaaccagt ttatccgtgg
     3901 cggcatcgcc ttcgccacgc ctccgggtac gccactggca ccgaaagccc aggaaggcaa
     3961 acacttcctg ttgcaggaaa gtgaaccgaa agagtggcgt gaatggggaa ctgcgcttcc
     4021 caaataatgc ccacctcgag cagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     4081 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     4141 gaggggtttt ttgctgaaag gaggaactat atccggataa ttcttgaaga cgaaagggcc
     4201 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4621 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4921 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6181 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6241 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     6301 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     6361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6421 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     6481 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     6541 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     6601 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     6661 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     6721 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     6781 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     6841 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     6901 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     6961 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     7021 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gag
//
