LOCUS       sequence-309829-        6020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 4 ring (rings 1347) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6020)
  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 6020)
  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: pBEL2042
FEATURES             Location/Qualifiers
     source          1..6020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        128..146
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             178..200
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3105..3123)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3119..3166
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3211..3229)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3297..3401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3402..3470,3471..4262)
                     /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(3620..3639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4433..5021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4922..4941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5175..5192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5207..5346
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5272..5291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5432..5454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5448..5639)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
       61 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tctcgatccc
      121 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt
      181 gtttaacttt aagaaggaga tatacatatg gctcttaaca caccacaaat tacgccgaca
      241 aaaaagataa cagtgagggc aatcggcgag gaactgccgc gtggtgatta ccaacgttgc
      301 ccgcaatgtg acatgctgtt tagcctgccc gagataaatt ctcatcaaag tgcctattgt
      361 ccgcgctgtc aggcaaaaat tcgtgacggg cgcgactggt cgctaacgcg cctggcggca
      421 atggccttca ccatgctgtt gcttatgccg tttgcctggg gcgaaccgct gttgcatatc
      481 tggctgttag gcattcgtat tgacgccaac gttatgcaag gcatctggca aatgaccaaa
      541 cagggcgatg cgataacggg gtcgatggtc tttttctgcg ttattggtgc cccgctcatt
      601 ctggtgacct ccatagctta tttatggttt ggtaaccgac tgggaatgaa tttacgtccg
      661 gtactgctga tgcttgagcg acttaaagag tgggtaatgc tcgatatcta cctggtcggc
      721 attggcgttg cttctataaa ggtacaggat tatgcccata tccaggctgg tgtcggcttg
      781 ttctcttttg tggcgttggt gattttaacg acggtgacgt tgtcacatct taatgtcgaa
      841 gaactgtggg agcgatttta tccgcagcgc cccgctacgc gtagggacga gaaacttcgt
      901 gtctgcctag ggtgccattt taccggctat ccagatcagc gtggtcgctg cccgcgttgc
      961 catatcccgc tacgcctgcg tcgccgtcat agtctgcaaa aatgctgggc ggcgctgtta
     1021 gcgtcaatcg ttttattgtt acctgccaac ctgttgccta tttctatcat ttatctgaac
     1081 ggaggacggc aggaagatac aattctttcc ggaattatgt cgctggcaag tagcaacatt
     1141 gcggttgcag gaatcgtgtt tatcgccagt attctggtcc cgtttactaa agtgatcgtc
     1201 atgttcactt tactgttgag cattcatttt aaatgccagc aaggtttacg cacacgcatt
     1261 ctgttactgc ggatggtgac ctggattggt cgctggtcga tgctcgacct gtttgtcata
     1321 tctttaacca tgtcgctgat taatcgcgat cagatcctcg cttttactat gggaccggct
     1381 gcgttttatt tcggcgcagc ggtaattttg actattcttg ctgtggaatg gctggacagc
     1441 cgcttacttt gggatgcaca tgagtcagga aacgcccgct tcgacgactg aagcgcagat
     1501 taaaaataaa cgccgtatct cacctttctg gctgctgccg ttcatcgcgc taatgattgc
     1561 cagttggctg atttgggaca gttatcagga ccggggtaat accgtcacca tcgactttat
     1621 gtcggcggat ggtattgttc caggccgtac gcctgttcgt tatcagggcg ttgaagtcgg
     1681 aacagtgcag gacatcagcc tcagcgacga tcttcgtaag attgaagtca aggtcagcat
     1741 caagtccgat atgaaagatg cgctgcgcga agagactcag ttctggctgg tgacgccaaa
     1801 agcatcgttg gcaggtgtct ccgggctgga cgccctcgtc ggtgggaact atatcggcat
     1861 gatgcccggg aaaggtaaag agcaggatca ctttgtcgca ctcgataccc aaccgaaata
     1921 tcggctggac aatggcggcg taataataaa actggaactg ccgagtgggg ccggattaac
     1981 cgccgactcg acgccgttaa tgtatcaggg gctggaagtc ggacagctga ctaaactgga
     2041 tttaaatcct ggtggtaaag tcacagggga aatgaccgtt gatcccagcg tcgttaccct
     2101 gcttcgtgaa aatacccgca tcgaattacg caacccgaaa ttatccctta gcgatgctaa
     2161 tctcagcgcc ctgctgaccg gcaaaacctt cgagctggtg cccggcgatg gcgagccacg
     2221 caaagaattc gttgttgtgc caggcgaaaa agcactgctg catgaacctg atgttctgac
     2281 gctgaccctg accgcgccgg aaagttacgg tattgatgcg ggtcagccgc tcattcttca
     2341 cggcgtgcag gtaggccagg ttatcgatcg taaactcacc agcaaaggcg tcacctttac
     2401 cgtcgccatc gagcctcagc atcgcgaact ggtaaaaggc gatagcaaat ttgtcgtcaa
     2461 cagccgtgtc gatgtgaagg tggggctgga tggcgttgag tttctcggtg ccagcgcctc
     2521 agaatggatc aatggcggga tccgtattct gccgggcgat aaaggtgaga tgaaagccag
     2581 ctatccactg tatgccaatt tggaaaaagc gctggagaac agccttagcg atttacccac
     2641 cacaggctta agcattattg ttgaagcgcc ggaagccggt tcgttaggca tcggtacgcc
     2701 tgtgctgttc cgtggtctgg aagtcggtac ggttacagga atgacgctgg ggacattgtc
     2761 agatcgcgtg atgattgcga tgcgcatcag taaacgctat caacacctgg tgcgtaacaa
     2821 ttccgtcttc tggttggcat cgggttacag tctggacttt ggtctgacgg gcggcgtagt
     2881 gaaaaccggc acctttaacc agtttatccg tggcggcatc gccttcgcca cgcctccggg
     2941 tacgccactg gcaccgaaag cccaggaagg caaacacttc ctgttgcagg aaagtgaacc
     3001 gaaagagtgg cgtgaatggg gaactgcgct tcccaaataa tgcccacctc gagcagatcc
     3061 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     3121 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     3181 tatatccgga taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3241 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3301 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3361 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3421 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3481 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3541 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3601 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     3661 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3721 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3781 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3841 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3901 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     3961 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4081 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4261 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4561 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5041 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5101 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5161 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     5221 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     5281 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     5341 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5401 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5461 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     5521 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     5581 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     5641 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     5701 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     5761 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     5821 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     5881 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     5941 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     6001 cttcacgttc gctcgcgtat
//
