LOCUS       sequence-309661-        6734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 6 ring (rings 123457) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6734)
  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 6734)
  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: pBEL2009
FEATURES             Location/Qualifiers
     source          1..6734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     217..236
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        217..235
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3908..3926)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3922..3969
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4014..4032)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4100..4204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4205..4273,4274..5065)
                     /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(4423..4442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5236..5824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5725..5744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5978..5995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6010..6149
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6075..6094)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6235..6257
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6251..6442)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
       61 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
      121 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
      181 gacccaacgc tgcccgagat ctcgatcccg cgaaattaat acgactcact atagggagac
      241 cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat atacatatgg
      301 ctcttaacac accacaaatt acgccgacaa aaaagataac agtgagggca atcggcgagg
      361 aactgccgcg tggtgattac caacgttgcc cgcaatgtga catgctgttt agcctgcccg
      421 agataaattc tcatcaaagt gcctattgtc cgcgctgtca ggcaaaaatt cgtgacgggc
      481 gcgactggtc gctaacgcgc ctggcggcaa tggccttcac catgctgttg cttatgccgt
      541 ttgcctgggg cgaaccgctg ttgcatatct ggctgttagg cattcgtatt gacgccaacg
      601 ttatgcaagg catctggcaa atgaccaaac agggcgatgc gataacgggg tcgatggtct
      661 ttttctgcgt tattggtgcc ccgctcattc tggtgacctc catagcttat ttatggtttg
      721 gtaaccgact gggaatgaat ttacgtccgg tactgctgat gcttgagcga cttaaagagt
      781 gggtaatgct cgatatctac ctggtcggca ttggcgttgc ttctataaag gtacaggatt
      841 atgcccatat ccaggctggt gtcggcttgt tctcttttgt ggcgttggtg attttaacga
      901 cggtgacgtt gtcacatctt aatgtcgaag aactgtggga gcgattttat ccgcagcgcc
      961 ccgctacgcg tagggacgag aaacttcgtg tctgcctagg gtgccatttt accggctatc
     1021 cagatcagcg tggtcgctgc ccgcgttgcc atatcccgct acgcctgcgt cgccgtcata
     1081 gtctgcaaaa atgctgggcg gcgctgttag cgtcaatcgt tttattgtta cctgccaacc
     1141 tgttgcctat ttctatcatt tatctgaacg gaggacggca ggaagataca attctttccg
     1201 gaattatgtc gctggcaagt agcaacattg cggttgcagg aatcgtgttt atcgccagta
     1261 ttctggtccc gtttactaaa gtgatcgtca tgttcacttt actgttgagc attcatttta
     1321 aatgccagca aggtttacgc acacgcattc tgttactgcg gatggtgacc tggattggtc
     1381 gctggtcgat gctcgacctg tttgtcatat ctttaaccat gtcgctgatt aatcgcgatc
     1441 agatcctcgc ttttactatg ggaccggctg cgttttattt cggcgcagcg gtaattttga
     1501 ctattcttgc tgtggaatgg ctggacagcc gcttactttg ggatgcacat gagtcaggaa
     1561 acgcccgctt cgacgactga agcgcagatt aaaaataaac gccgtatctc acctttctgg
     1621 ctgctgccgt tcatcgcgct aatgattgcc agttggctga tttgggacag ttatcaggac
     1681 cggggtaata ccgtcaccat cgactttatg tcggcggatg gtattgttcc aggccgtacg
     1741 cctgttcgtt atcagggcgt tgaagtcgga acagtgcagg acatcagcct cagcgacgat
     1801 cttcgtaaga ttgaagtcaa ggtcagcatc aagtccgata tgaaagatgc gctgcgcgaa
     1861 gagactcagt tctggctggt gacgccaaaa gcatcgttgg caggtgtctc cgggctggac
     1921 gccctcgtcg gtgggaacta tatcggcatg atgccgggta aaggtaaaga gcaggatcac
     1981 tttgtcgcac tcgataccca accgaaatat cggctggaca atggcgatct gatgatccac
     2041 ctgcaagccc ccgatctcgg ttcgctgaac agcggttcat tggtctattt ccgcaagatc
     2101 ccggtgggaa aagtctacga ctatgccatc aatcccaaca agcaaggcgt ggtgattgat
     2161 gtcctgatcg agcggcgttt taccgatctg gtgaaaaaag gtagccgttt ctggaacgtt
     2221 tccggcgttg atgccaacgt cagtatcagt ggcgcgaagg tgaaactgga aagcctggcg
     2281 gcactggtta acggtgcgat tgccttcgat tcaccagaag agtcgaaacc tgccgaggcg
     2341 gaagatacct ttggtctgta tgaagatctg gcccacagcc agcgtggcgt aataataaaa
     2401 ctggaactgc cgagtggggc cggattaacc gccgactcga cgccgttaat gtatcagggg
     2461 ctggaagtcg gacagctgac taaactggat ttaaatcctg gtggtaaagt cacaggggaa
     2521 atgaccgttg atcccagcgt cgttaccctg cttcgtgaaa atacccgcat cgaattacgc
     2581 aacccgaaat tatcccttag cgatgctaat ctcagcgccc tgctgaccgg caaaaccttc
     2641 gagctggtgc ccggcgatgg cgagccacgc aaagagttcg ttgttgtgcc aggcgaaaaa
     2701 gcactgctgc atgaacctga tgttctgacg ctgaccctga ccgcgccgga aagttacggt
     2761 attgatgcgg gtcagccgct cattcttcac ggcgtgcagg taggccaggt tatcgatcgt
     2821 aaactcacca gcaaaggcgt cacctttacc gtcgccatcg agcctcagca tcgcgaactg
     2881 gtaaaaggcg atagcaaatt tgtcgtcaac agccgtgtcg atgtgaaggt ggggctggat
     2941 ggcgttgagt ttctcggtgc cagcgcctca gaatggatca atggcgggat acgtattctg
     3001 ccgggcgata aaggtgagat gaaagccagc tatccactgt atgccaattt ggaaaaagcg
     3061 ctggagaaca gccttagcga tttacccacc acaaccgtga gtttgagtgc agagacgctg
     3121 ccggatgtgc aggcaggatc ggtagtgctg taccgtaaat ttgaagttgg tgaagtgatt
     3181 accgtgcgtc cgcgagctaa cgcgtttgat atcgatctgc atattaagcc ggagtatcgc
     3241 aaccttctga ccagcaatag cgtgttctgg gcagaaggcg gggcgaaagt tcagctgaat
     3301 ggtagtggtc tgaccgtaca ggcatccccg ctctccagag cattaaaggg agccattagc
     3361 ttcgacaacc tcagcggtgc cagcgccagt cagcgtaaag gcgataagcg tattctgtat
     3421 gcttccgaaa cagcggcccg tgcggttggc ttaagcatta ttgttgaagc gccggaagcc
     3481 ggttcgttag gcatcggtac gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca
     3541 ggaatgacgc tggggacatt gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc
     3601 tatcaacacc tggtgcgtaa caattccgtc ttctggttgg catcgggtta cagtctggac
     3661 tttggtctga cgggcggcgt agtgaaaacc ggcaccttta accagtttat ccgtggcggc
     3721 atcgccttcg ccacgcctcc gggtacgcca ctggcaccga aagcccagga aggcaaacac
     3781 ttcctgttgc aggaaagtga accgaaagag tggcgtgaat ggggaactgc gcttcccaaa
     3841 taatgcccac ctcgagcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     3901 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3961 ggttttttgc tgaaaggagg aactatatcc ggataattct tgaagacgaa agggcctcgt
     4021 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4081 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4141 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4201 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4261 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4321 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4381 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4441 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4501 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4561 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4621 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4681 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4741 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4801 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4861 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4921 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4981 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5041 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5341 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5521 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5581 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5641 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5701 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5761 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5821 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5881 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5941 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6001 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     6061 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     6121 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     6181 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6241 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     6301 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     6361 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     6421 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     6481 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     6541 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     6601 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     6661 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     6721 gcgtttccag actt
//
