LOCUS       sequence-310596-        7123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB operon with LetA and LetB genes 
            deoverlapped.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7123)
  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 7123)
  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: pBEL2071
FEATURES             Location/Qualifiers
     source          1..7123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     285..302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    317..456
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(382..401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     542..564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(558..749)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1258..1277
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1258..1276
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1308..1330
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5338..5356)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5352..5399
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5444..5462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5530..5634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5635..5703,5704..6495)
                     /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(5853..5872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6666..7123,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      301 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
      361 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
      421 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
      481 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
      541 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
      601 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
      661 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
      721 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
      781 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
      841 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
      901 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
      961 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     1021 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     1081 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     1141 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     1201 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa
     1261 tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt
     1321 aagaaggaga tatacatatg gctcttaaca caccacaaat tacgccgaca aaaaagataa
     1381 cagtgagggc aatcggcgag gaactgccgc gtggtgatta ccaacgttgc ccgcaatgtg
     1441 acatgctgtt tagcctgccc gagataaatt ctcatcaaag tgcctattgt ccgcgctgtc
     1501 aggcaaaaat tcgtgacggg cgcgactggt cgctaacgcg cctggcggca atggccttca
     1561 ccatgctgtt gcttatgccg tttgcctggg gcgaaccgct gttgcatatc tggctgttag
     1621 gcattcgtat tgacgccaac gttatgcaag gcatctggca aatgaccaaa cagggcgatg
     1681 cgataacggg gtcgatggtc tttttctgcg ttattggtgc cccgctcatt ctggtgacct
     1741 ccatagctta tttatggttt ggtaaccgac tgggaatgaa tttacgtccg gtactgctga
     1801 tgcttgagcg acttaaagag tgggtaatgc tcgatatcta cctggtcggc attggcgttg
     1861 cttctataaa ggtacaggat tatgcccata tccaggctgg tgtcggcttg ttctcttttg
     1921 tggcgttggt gattttaacg acggtgacgt tgtcacatct taatgtcgaa gaactgtggg
     1981 agcgatttta tccgcagcgc cccgctacgc gtagggacga gaaacttcgt gtctgcctag
     2041 ggtgccattt taccggctat ccagatcagc gtggtcgctg cccgcgttgc catatcccgc
     2101 tacgcctgcg tcgccgtcat agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg
     2161 ttttattgtt acctgccaac ctgttgccta tttctatcat ttatctgaac ggaggacggc
     2221 aggaagatac aattctttcc ggaattatgt cgctggcaag tagcaacatt gcggttgcag
     2281 gaatcgtgtt tatcgccagt attctggtcc cgtttactaa agtgatcgtc atgttcactt
     2341 tactgttgag cattcatttt aaatgccagc aaggtttacg cacacgcatt ctgttactgc
     2401 ggatggtgac ctggattggt cgctggtcga tgctcgacct gtttgtcata tctttaacca
     2461 tgtcgctgat taatcgcgat cagatcctcg cttttactat gggaccggct gcgttttatt
     2521 tcggcgcagc ggtaattttg actattcttg ctgtggaatg gctggacagc cgcttacttt
     2581 gggatgcaca cgaaagcggc aatgcgcgtt ttgatgattg atttcaggag aaccgacgca
     2641 tgagtcagga aacgcccgct tcgacgactg aagcgcagat taaaaataaa cgccgtatct
     2701 cacctttctg gctgctgccg ttcatcgcgc taatgattgc cagttggctg atttgggaca
     2761 gttatcagga ccggggtaat accgtcacca tcgactttat gtcggcggat ggtattgttc
     2821 caggccgtac gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag gacatcagcc
     2881 tcagcgacga tcttcgtaag attgaagtca aggtcagcat caagtccgat atgaaagatg
     2941 cgctgcgcga agagactcag ttctggctgg tgacgccaaa agcatcgttg gcaggtgtct
     3001 ccgggctgga cgccctcgtc ggtgggaact atatcggcat gatgcccggg aaaggtaaag
     3061 agcaggatca ctttgtcgca ctcgataccc aaccgaaata tcggctggac aatggcgatc
     3121 tgatgatcca cctgcaagcc cccgatctcg gttcgctgaa cagcggttca ttggtctatt
     3181 tccgcaagat cccggtggga aaagtctacg actatgccat caatcccaac aagcaaggcg
     3241 tggtgattga tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa ggtagccgtt
     3301 tctggaacgt ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag gtgaaactgg
     3361 aaagcttggc ggcactggtt aacggtgcga ttgccttcga ttcaccagaa gagtcgaaac
     3421 ctgccgaggc ggaagatacc tttggtctgt atgaagatct ggcccacagc cagcgtggcg
     3481 taataataaa actggaactg ccgagtgggg ccggattaac cgccgactcg acgccgttaa
     3541 tgtatcaggg gctggaagtc ggacagctga ctaaactgga tttaaatcct ggtggtaaag
     3601 tcacagggga aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa aatacccgca
     3661 tcgaattacg caacccgaaa ttatccctta gcgatgctaa tctcagcgcc ctgctgaccg
     3721 gcaaaacctt cgagctggtg cccggcgatg gcgagccacg caaagaattc gttgttgtgc
     3781 caggcgaaaa agcactgctg catgaacctg atgttctgac gctgaccctg accgcgccgg
     3841 aaagttacgg tattgatgcg ggtcagccgc tcattcttca cggcgtgcag gtaggccagg
     3901 ttatcgatcg taaactcacc agcaaaggcg tcacctttac cgtcgccatc gagcctcagc
     3961 atcgcgaact ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc gatgtgaagg
     4021 tggggctgga tggcgttgag tttctcggtg ccagcgcctc agaatggatc aatggcggga
     4081 tccgtattct gccgggcgat aaaggtgaga tgaaagccag ctatccactg tatgccaatt
     4141 tggaaaaagc gctggagaac agccttagcg atttacccac cacaaccgtg agtttgagtg
     4201 cagagacgct gccggatgtg caggcaggat cggtagtgct gtaccgtaaa tttgaagttg
     4261 gtgaagtgat taccgtgcgt ccgcgagcta acgcgtttga catcgatctg catattaagc
     4321 cggagtatcg caaccttctg accagcaata gcgtgttctg ggcagaaggc ggggcgaaag
     4381 ttcagctgaa tggtagtggt ctgaccgtac aggcatcccc gctctccaga gctctaaagg
     4441 gagccattag cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa ggcgataagc
     4501 gtattctgta tgcttccgaa acagcggccc gtgcggttgg cgggcagatt acgcttcacg
     4561 ctttcgatgc cggaaaactg gcggtcggga tgccaattcg ctatctcggt attgacatcg
     4621 ggcaaatcca gacgctggat ctgattaccg cacgcaatga agtacaggca aaggcggtac
     4681 tttacccgga atatgtccag acctttgctc gcggtggtac gcgcttctca gtggtcacac
     4741 cgcaaatttc ggcagctggc gttgagcatc ttgatactat cctccagccg tatatcaacg
     4801 tcgaaccagg ccggggcaat cctcgccgcg actttgaatt acaagaggcc accattactg
     4861 attcgcggta cctggatggc ttaagcatta ttgttgaagc gccggaagcc ggttcgttag
     4921 gcatcggtac gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca ggaatgacgc
     4981 tggggacatt gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc tatcaacacc
     5041 tggtgcgtaa caattccgtc ttctggttgg catcgggtta cagtctggac tttggtctga
     5101 cgggcggcgt agtgaaaacc ggcaccttta accagtttat ccgtggcggc atcgccttcg
     5161 ccacgcctcc gggtacgcca ctggcaccga aagcccagga aggcaaacac ttcctgttgc
     5221 aggaaagtga accgaaagag tggcgtgaat ggggaactgc gcttcccaaa taatgcccac
     5281 ctcgagcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     5341 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     5401 tgaaaggagg aactatatcc ggataattct tgaagacgaa agggcctcgt gatacgccta
     5461 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5521 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5581 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5641 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5701 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5761 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5821 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5881 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5941 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6001 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6061 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6121 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6181 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6241 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6301 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6361 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6421 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6481 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6541 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6601 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6661 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6721 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6781 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6841 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6901 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6961 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7021 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7081 gaagggagaa aggcggacag gtatccggta agcggcaggg tcg
//
