LOCUS       sequence-309763-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB mutant L359N.
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: pBEL1862
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     616..635
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        616..634
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             666..688
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4646..4664)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4660..4707
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4752..4770)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4838..4942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4943..5011,5012..5803)
                     /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(5161..5180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5974..6562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6463..6482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6716..6733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6748..6887
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6813..6832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6973..6995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(6989..7073,1..107))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
       61 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
      121 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
      181 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
      241 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
      301 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
      361 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
      421 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
      481 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
      541 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatc
      601 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
      661 ataattttgt ttaactttaa gaaggagata tacatatggc tcttaacaca ccacaaatta
      721 cgccgacaaa aaagataaca gtgagggcaa tcggcgagga actgccgcgt ggtgattacc
      781 aacgttgccc gcaatgtgac atgctgttta gcctgcccga gataaattct catcaaagtg
      841 cctattgtcc gcgctgtcag gcaaaaattc gtgacgggcg cgactggtcg ctaacgcgcc
      901 tggcggcaat ggccttcacc atgctgttgc ttatgccgtt tgcctggggc gaaccgctgt
      961 tgcatatctg gctgttaggc attcgtattg acgccaacgt tatgcaaggc atctggcaaa
     1021 tgaccaaaca gggcgatgcg ataacggggt cgatggtctt tttctgcgtt attggtgccc
     1081 cgctcattct ggtgacctcc atagcttatt tatggtttgg taaccgactg ggaatgaatt
     1141 tacgtccggt actgctgatg cttgagcgac ttaaagagtg ggtaatgctc gatatctacc
     1201 tggtcggcat tggcgttgct tctataaagg tacaggatta tgcccatatc caggctggtg
     1261 tcggcttgtt ctcttttgtg gcgttggtga ttttaacgac ggtgacgttg tcacatctta
     1321 atgtcgaaga actgtgggag cgattttatc cgcagcgccc cgctacgcgt agggacgaga
     1381 aacttcgtgt ctgcctaggg tgccatttta ccggctatcc agatcagcgt ggtcgctgcc
     1441 cgcgttgcca tatcccgcta cgcctgcgtc gccgtcatag tctgcaaaaa tgctgggcgg
     1501 cgctgttagc gtcaatcgtt ttattgttac ctgccaacct gttgcctatt tctatcattt
     1561 atctgaacgg aggacggcag gaagatacaa ttctttccgg aattatgtcg ctggcaagta
     1621 gcaacattgc ggttgcagga atcgtgttta tcgccagtat tctggtcccg tttactaaag
     1681 tgatcgtcat gttcacttta ctgttgagca ttcattttaa atgccagcaa ggtttacgca
     1741 cacgcattct gttactgcgg atggtgacct ggattggtcg ctggtcgatg ctcgacctgt
     1801 ttgtcatatc tttaaccatg tcgctgatta atcgcgatca gatcctcgct tttactatgg
     1861 gaccggctgc gttttatttc ggcgcagcgg taattttgac tattcttgct gtggaatggc
     1921 tggacagccg cttactttgg gatgcacatg agtcaggaaa cgcccgcttc gacgactgaa
     1981 gcgcagatta aaaataaacg ccgtatctca cctttctggc tgctgccgtt catcgcgcta
     2041 atgattgcca gttggctgat ttgggacagt tatcaggacc ggggtaatac cgtcaccatc
     2101 gactttatgt cggcggatgg tattgttcca ggccgtacgc ctgttcgtta tcagggcgtt
     2161 gaagtcggaa cagtgcagga catcagcctc agcgacgatc ttcgtaagat tgaagtcaag
     2221 gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag agactcagtt ctggctggtg
     2281 acgccaaaag catcgttggc aggtgtctcc gggctggacg ccctcgtcgg tgggaactat
     2341 atcggcatga tgccgggtaa aggtaaagag caggatcact ttgtcgcact cgatacccaa
     2401 ccgaaatatc ggctggacaa tggcgatctg atgatccacc tgcaagcccc cgatctcggt
     2461 tcgctgaaca gcggttcatt ggtctatttc cgcaagatcc cggtgggaaa agtctacgac
     2521 tatgccatca atcccaacaa gcaaggcgtg gtgattgatg tcctgatcga gcggcgtttt
     2581 accgatctgg tgaaaaaagg tagccgtttc tggaacgttt ccggcgttga tgccaacgtc
     2641 agtatcagtg gcgcgaaggt gaaactggaa agcctggcgg cactggttaa cggtgcgatt
     2701 gccttcgatt caccagaaga gtcgaaacct gccgaggcgg aagatacctt tggtctgtat
     2761 gaagatctgg cccacagcca gcgtggcgta ataataaaac tggaactgcc gagtggggcc
     2821 ggattaaccg ccgactcgac gccgttaatg tatcaggggc tggaagtcgg acagctgact
     2881 aaactggatt taaatcctgg tggtaaagtc acaggggaaa tgaccgttga tcccagcgtc
     2941 gttaccctgc ttcgtgaaaa tacccgcatc gaattacgca acccgaaatt atcccttagc
     3001 gatgctaatc tcagcgccct gaataccggc aaaaccttcg agctggtgcc cggcgatggc
     3061 gagccacgca aagagttcgt tgttgtgcca ggcgaaaaag cactgctgca tgaacctgat
     3121 gttctgacgc tgaccctgac cgcgccggaa agttacggta ttgatgcggg tcagccgctc
     3181 attcttcacg gcgtgcaggt aggccaggtt atcgatcgta aactcaccag caaaggcgtc
     3241 acctttaccg tcgccatcga gcctcagcat cgcgaactgg taaaaggcga tagcaaattt
     3301 gtcgtcaaca gccgtgtcga tgtgaaggtg gggctggatg gcgttgagtt tctcggtgcc
     3361 agcgcctcag aatggatcaa tggcgggata cgtattctgc cgggcgataa aggtgagatg
     3421 aaagccagct atccactgta tgccaatttg gaaaaagcgc tggagaacag ccttagcgat
     3481 ttacccacca caaccgtgag tttgagtgca gagacgctgc cggatgtgca ggcaggatcg
     3541 gtagtgctgt accgtaaatt tgaagttggt gaagtgatta ccgtgcgtcc gcgagctaac
     3601 gcgtttgaca tcgatctgca tattaagccg gagtatcgca accttctgac cagcaatagc
     3661 gtgttctggg cagaaggcgg ggcgaaagtt cagctgaatg gtagtggtct gaccgtacag
     3721 gcatccccgc tctccagagc tctaaaggga gccattagct tcgacaacct cagcggtgcc
     3781 agcgccagtc agcgtaaagg cgataagcgt attctgtatg cttccgaaac agcggcccgt
     3841 gcggttggcg ggcagattac gcttcacgct ttcgatgccg gaaaactggc ggtcgggatg
     3901 ccaattcgct atctcggtat tgacatcggg caaatccaga cgctggatct gattaccgca
     3961 cgcaatgaag tacaggcaaa ggcggtactt tacccggaat atgtccagac ctttgctcgc
     4021 ggtggtacgc gcttctcagt ggtcacaccg caaatttcgg cagctggcgt tgagcatctt
     4081 gatactatcc tccagccgta tatcaacgtc gaaccaggcc ggggcaatcc tcgccgcgac
     4141 tttgaattac aagaggccac cattactgat tcgcggtacc tggatggctt aagcattatt
     4201 gttgaagcgc cggaagccgg ttcgttaggc atcggtacgc ctgtgctgtt ccgtggtctg
     4261 gaagtcggta cggttacagg aatgacgctg gggacattgt cagatcgcgt gatgattgcg
     4321 atgcgcatca gtaaacgcta tcaacacctg gtgcgtaaca attccgtctt ctggttggca
     4381 tcgggttaca gtctggactt tggtctgacg ggcggcgtag tgaaaaccgg cacctttaac
     4441 cagtttatcc gtggcggcat cgccttcgcc acgcctccgg gtacgccact ggcaccgaaa
     4501 gcccaggaag gcaaacactt cctgttgcag gaaagtgaac cgaaagagtg gcgtgaatgg
     4561 ggaactgcgc ttcccaaata atgcccacct cgagcagatc cggctgctaa caaagcccga
     4621 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     4681 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ataattcttg
     4741 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4801 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4861 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4921 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4981 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5041 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5101 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5161 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     5221 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5281 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5341 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5401 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5461 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     5521 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5581 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5641 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5701 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5761 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5821 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5881 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5941 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6061 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6121 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6181 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6301 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6361 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6421 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6481 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6541 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6601 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6661 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6721 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     6781 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6841 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     6901 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6961 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     7021 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atg
//
