LOCUS       sequence-309658-        6377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 5 ring (rings 12347) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  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 6377)
  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: pBEL2008
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             262..284
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3546..3564)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3560..3607
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3652..3670)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3738..3842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3843..3911,3912..4703)
                     /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(4061..4080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4874..5462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5363..5382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5616..5633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5648..5787
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5713..5732)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5873..5895
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5889..6080)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
       61 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
      121 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
      181 aacgctgccc gagatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
      241 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatggctctt
      301 aacacaccac aaattacgcc gacaaaaaag ataacagtga gggcaatcgg cgaggaactg
      361 ccgcgtggtg attaccaacg ttgcccgcaa tgtgacatgc tgtttagcct gcccgagata
      421 aattctcatc aaagtgccta ttgtccgcgc tgtcaggcaa aaattcgtga cgggcgcgac
      481 tggtcgctaa cgcgcctggc ggcaatggcc ttcaccatgc tgttgcttat gccgtttgcc
      541 tggggcgaac cgctgttgca tatctggctg ttaggcattc gtattgacgc caacgttatg
      601 caaggcatct ggcaaatgac caaacagggc gatgcgataa cggggtcgat ggtctttttc
      661 tgcgttattg gtgccccgct cattctggtg acctccatag cttatttatg gtttggtaac
      721 cgactgggaa tgaatttacg tccggtactg ctgatgcttg agcgacttaa agagtgggta
      781 atgctcgata tctacctggt cggcattggc gttgcttcta taaaggtaca ggattatgcc
      841 catatccagg ctggtgtcgg cttgttctct tttgtggcgt tggtgatttt aacgacggtg
      901 acgttgtcac atcttaatgt cgaagaactg tgggagcgat tttatccgca gcgccccgct
      961 acgcgtaggg acgagaaact tcgtgtctgc ctagggtgcc attttaccgg ctatccagat
     1021 cagcgtggtc gctgcccgcg ttgccatatc ccgctacgcc tgcgtcgccg tcatagtctg
     1081 caaaaatgct gggcggcgct gttagcgtca atcgttttat tgttacctgc caacctgttg
     1141 cctatttcta tcatttatct gaacggagga cggcaggaag atacaattct ttccggaatt
     1201 atgtcgctgg caagtagcaa cattgcggtt gcaggaatcg tgtttatcgc cagtattctg
     1261 gtcccgttta ctaaagtgat cgtcatgttc actttactgt tgagcattca ttttaaatgc
     1321 cagcaaggtt tacgcacacg cattctgtta ctgcggatgg tgacctggat tggtcgctgg
     1381 tcgatgctcg acctgtttgt catatcttta accatgtcgc tgattaatcg cgatcagatc
     1441 ctcgctttta ctatgggacc ggctgcgttt tatttcggcg cagcggtaat tttgactatt
     1501 cttgctgtgg aatggctgga cagccgctta ctttgggatg cacatgagtc aggaaacgcc
     1561 cgcttcgacg actgaagcgc agattaaaaa taaacgccgt atctcacctt tctggctgct
     1621 gccgttcatc gcgctaatga ttgccagttg gctgatttgg gacagttatc aggaccgggg
     1681 taataccgtc accatcgact ttatgtcggc ggatggtatt gttccaggcc gtacgcctgt
     1741 tcgttatcag ggcgttgaag tcggaacagt gcaggacatc agcctcagcg acgatcttcg
     1801 taagattgaa gtcaaggtca gcatcaagtc cgatatgaaa gatgcgctgc gcgaagagac
     1861 tcagttctgg ctggtgacgc caaaagcatc gttggcaggt gtctccgggc tggacgccct
     1921 cgtcggtggg aactatatcg gcatgatgcc gggtaaaggt aaagagcagg atcactttgt
     1981 cgcactcgat acccaaccga aatatcggct ggacaatggc gatctgatga tccacctgca
     2041 agcccccgat ctcggttcgc tgaacagcgg ttcattggtc tatttccgca agatcccggt
     2101 gggaaaagtc tacgactatg ccatcaatcc caacaagcaa ggcgtggtga ttgatgtcct
     2161 gatcgagcgg cgttttaccg atctggtgaa aaaaggtagc cgtttctgga acgtttccgg
     2221 cgttgatgcc aacgtcagta tcagtggcgc gaaggtgaaa ctggaaagcc tggcggcact
     2281 ggttaacggt gcgattgcct tcgattcacc agaagagtcg aaacctgccg aggcggaaga
     2341 tacctttggt ctgtatgaag atctggccca cagccagcgt ggcgtaataa taaaactgga
     2401 actgccgagt ggggccggat taaccgccga ctcgacgccg ttaatgtatc aggggctgga
     2461 agtcggacag ctgactaaac tggatttaaa tcctggtggt aaagtcacag gggaaatgac
     2521 cgttgatccc agcgtcgtta ccctgcttcg tgaaaatacc cgcatcgaat tacgcaaccc
     2581 gaaattatcc cttagcgatg ctaatctcag cgccctgctg accggcaaaa ccttcgagct
     2641 ggtgcccggc gatggcgagc cacgcaaaga gttcgttgtt gtgccaggcg aaaaagcact
     2701 gctgcatgaa cctgatgttc tgacgctgac cctgaccgcg ccggaaagtt acggtattga
     2761 tgcgggtcag ccgctcattc ttcacggcgt gcaggtaggc caggttatcg atcgtaaact
     2821 caccagcaaa ggcgtcacct ttaccgtcgc catcgagcct cagcatcgcg aactggtaaa
     2881 aggcgatagc aaatttgtcg tcaacagccg tgtcgatgtg aaggtggggc tggatggcgt
     2941 tgagtttctc ggtgccagcg cctcagaatg gatcaatggc gggatacgta ttctgccggg
     3001 cgataaaggt gagatgaaag ccagctatcc actgtatgcc aatttggaaa aagcgctgga
     3061 gaacagcctt agcgatttac ccaccggctt aagcattatt gttgaagcgc cggaagccgg
     3121 ttcgttaggc atcggtacgc ctgtgctgtt ccgtggtctg gaagtcggta cggttacagg
     3181 aatgacgctg gggacattgt cagatcgcgt gatgattgcg atgcgcatca gtaaacgcta
     3241 tcaacacctg gtgcgtaaca attccgtctt ctggttggca tcgggttaca gtctggactt
     3301 tggtctgacg ggcggcgtag tgaaaaccgg cacctttaac cagtttatcc gtggcggcat
     3361 cgccttcgcc acgcctccgg gtacgccact ggcaccgaaa gcccaggaag gcaaacactt
     3421 cctgttgcag gaaagtgaac cgaaagagtg gcgtgaatgg ggaactgcgc ttcccaaata
     3481 atgcccacct cgagcagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     3541 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3601 ttttttgctg aaaggaggaa ctatatccgg ataattcttg aagacgaaag ggcctcgtga
     3661 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3901 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3961 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4021 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4081 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4141 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4201 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4261 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4321 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4381 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4441 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4501 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4561 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4621 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4681 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4741 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4801 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4861 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4921 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4981 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5041 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5101 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5161 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5221 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5281 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5341 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5401 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5461 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5521 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5581 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5641 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     5701 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     5761 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     5821 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5881 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     5941 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     6001 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     6061 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     6121 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     6181 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     6241 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     6301 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     6361 gtttccagac tttacga
//
