LOCUS       sequence-309718-        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB PLL1 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  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 7028)
  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: pBEL1886
FEATURES             Location/Qualifiers
     source          1..7028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..165
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        146..164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             196..218
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4131..4149)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4145..4192
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4237..4255)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4323..4427
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4428..4496,4497..5288)
                     /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(4646..4665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5459..6047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5948..5967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6201..6218
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6233..6372
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6298..6317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6458..6480
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6474..6665)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
       61 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
      121 gcccgagatc tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt
      181 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggc tcttaacaca
      241 ccacaaatta cgccgacaaa aaagataaca gtgagggcaa tcggcgagga actgccgcgt
      301 ggtgattacc aacgttgccc gcaatgtgac atgctgttta gcctgcccga gataaattct
      361 catcaaagtg cctattgtcc gcgctgtcag gcaaaaattc gtgacgggcg cgactggtcg
      421 ctaacgcgcc tggcggcaat ggccttcacc atgctgttgc ttatgccgtt tgcctggggc
      481 gaaccgctgt tgcatatctg gctgttaggc attcgtattg acgccaacgt tatgcaaggc
      541 atctggcaaa tgaccaaaca gggcgatgcg ataacggggt cgatggtctt tttctgcgtt
      601 attggtgccc cgctcattct ggtgacctcc atagcttatt tatggtttgg taaccgactg
      661 ggaatgaatt tacgtccggt actgctgatg cttgagcgac ttaaagagtg ggtaatgctc
      721 gatatctacc tggtcggcat tggcgttgct tctataaagg tacaggatta tgcccatatc
      781 caggctggtg tcggcttgtt ctcttttgtg gcgttggtga ttttaacgac ggtgacgttg
      841 tcacatctta atgtcgaaga actgtgggag cgattttatc cgcagcgccc cgctacgcgt
      901 agggacgaga aacttcgtgt ctgcctaggg tgccatttta ccggctatcc agatcagcgt
      961 ggtcgctgcc cgcgttgcca tatcccgcta cgcctgcgtc gccgtcatag tctgcaaaaa
     1021 tgctgggcgg cgctgttagc gtcaatcgtt ttattgttac ctgccaacct gttgcctatt
     1081 tctatcattt atctgaacgg aggacggcag gaagatacaa ttctttccgg aattatgtcg
     1141 ctggcaagta gcaacattgc ggttgcagga atcgtgttta tcgccagtat tctggtcccg
     1201 tttactaaag tgatcgtcat gttcacttta ctgttgagca ttcattttaa atgccagcaa
     1261 ggtttacgca cacgcattct gttactgcgg atggtgacct ggattggtcg ctggtcgatg
     1321 ctcgacctgt ttgtcatatc tttaaccatg tcgctgatta atcgcgatca gatcctcgct
     1381 tttactatgg gaccggctgc gttttatttc ggcgcagcgg taattttgac tattcttgct
     1441 gtggaatggc tggacagccg cttactttgg gatgcacatg agtcaggaaa cgcccgcttc
     1501 gacgactgaa gcgcagatta aaaataaacg ccgtatctca cctttctggc tgctgccgtt
     1561 catcgcgcta atgattgcca gttggctgat ttgggacagt tatcaggacc ggggtaatac
     1621 cgtcaccatc gactttatgt cggcggatgg tattgttcca ggccgtacgc ctgttcgtta
     1681 tcagggcgtt gaagtcggaa cagtgcagga catcagcctc agcgacgatc ttcgtaagat
     1741 tgaagtcaag gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag agactcagtt
     1801 ctggctggtg acgggaggga actatatcgg catgatgccc gggaaaggta aagagcagga
     1861 tcactttgtc gcactcgata cccaaccgaa atatcggctg gacaatggcg atctgatgat
     1921 ccacctgcaa gcccccgatc tcggttcgct gaacagcggt tcattggtct atttccgcaa
     1981 gatcccggtg ggaaaagtct acgactatgc catcaatccc aacaagcaag gcgtggtgat
     2041 tgatgtcctg atcgagcggc gttttaccga tctggtgaaa aaaggtagcc gtttctggaa
     2101 cgtttccggc gttgatgcca acgtcagtat cagtggcgcg aaggtgaaac tggaaagctt
     2161 ggcggcactg gttaacggtg cgattgcctt cgattcacca gaagagtcga aacctgccga
     2221 ggcggaagat acctttggtc tgtatgaaga tctggcccac agccagcgtg gcgtaataat
     2281 aaaactggaa ctgccgagtg gggccggatt aaccgccgac tcgacgccgt taatgtatca
     2341 ggggctggaa gtcggacagc tgactaaact ggatttaaat cctggtggta aagtcacagg
     2401 ggaaatgacc gttgatccca gcgtcgttac cctgcttcgt gaaaataccc gcatcgaatt
     2461 acgcaacccg aaattatccc ttagcgatgc taatctcagc gccctgctga ccggcaaaac
     2521 cttcgagctg gtgcccggcg atggcgagcc acgcaaagaa ttcgttgttg tgccaggcga
     2581 aaaagcactg ctgcatgaac ctgatgttct gacgctgacc ctgaccgcgc cggaaagtta
     2641 cggtattgat gcgggtcagc cgctcattct tcacggcgtg caggtaggcc aggttatcga
     2701 tcgtaaactc accagcaaag gcgtcacctt taccgtcgcc atcgagcctc agcatcgcga
     2761 actggtaaaa ggcgatagca aatttgtcgt caacagccgt gtcgatgtga aggtggggct
     2821 ggatggcgtt gagtttctcg gtgccagcgc ctcagaatgg atcaatggcg ggatccgtat
     2881 tctgccgggc gataaaggtg agatgaaagc cagctatcca ctgtatgcca atttggaaaa
     2941 agcgctggag aacagcctta gcgatttacc caccacaacc gtgagtttga gtgcagagac
     3001 gctgccggat gtgcaggcag gatcggtagt gctgtaccgt aaatttgaag ttggtgaagt
     3061 gattaccgtg cgtccgcgag ctaacgcgtt tgacatcgat ctgcatatta agccggagta
     3121 tcgcaacctt ctgaccagca atagcgtgtt ctgggcagaa ggcggggcga aagttcagct
     3181 gaatggtagt ggtctgaccg tacaggcatc cccgctctcc agagctctaa agggagccat
     3241 tagcttcgac aacctcagcg gtgccagcgc cagtcagcgt aaaggcgata agcgtattct
     3301 gtatgcttcc gaaacagcgg cccgtgcggt tggcgggcag attacgcttc acgctttcga
     3361 tgccggaaaa ctggcggtcg ggatgccaat tcgctatctc ggtattgaca tcgggcaaat
     3421 ccagacgctg gatctgatta ccgcacgcaa tgaagtacag gcaaaggcgg tactttaccc
     3481 ggaatatgtc cagacctttg ctcgcggtgg tacgcgcttc tcagtggtca caccgcaaat
     3541 ttcggcagct ggcgttgagc atcttgatac tatcctccag ccgtatatca acgtcgaacc
     3601 aggccggggc aatcctcgcc gcgactttga attacaagag gccaccatta ctgattcgcg
     3661 gtacctggat ggcttaagca ttattgttga agcgccggaa gccggttcgt taggcatcgg
     3721 tacgcctgtg ctgttccgtg gtctggaagt cggtacggtt acaggaatga cgctggggac
     3781 attgtcagat cgcgtgatga ttgcgatgcg catcagtaaa cgctatcaac acctggtgcg
     3841 taacaattcc gtcttctggt tggcatcggg ttacagtctg gactttggtc tgacgggcgg
     3901 cgtagtgaaa accggcacct ttaaccagtt tatccgtggc ggcatcgcct tcgccacgcc
     3961 tccgggtacg ccactggcac cgaaagccca ggaaggcaaa cacttcctgt tgcaggaaag
     4021 tgaaccgaaa gagtggcgtg aatggggaac tgcgcttccc aaataatgcc cacctcgagc
     4081 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     4141 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     4201 aggaactata tccggataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     4261 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4321 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4381 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4441 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4501 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4561 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4621 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     4681 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4741 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4801 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4861 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4921 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     4981 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5041 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5101 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5161 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5221 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5281 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5341 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5401 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5461 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5521 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5581 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5641 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5701 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5761 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5821 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5881 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5941 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6001 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6061 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6121 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6181 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     6241 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     6301 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     6361 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     6421 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     6481 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     6541 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     6601 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     6661 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     6721 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     6781 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     6841 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     6901 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     6961 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     7021 cagtcgct
//
