LOCUS       sequence-270802-        6008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 4 ring (rings 1237) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6008)
  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 6008)
  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: pBEL2041
FEATURES             Location/Qualifiers
     source          1..6008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3027..3045)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3041..3088
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3133..3151)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3219..3323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3324..3392,3393..4184)
                     /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(3542..3561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4355..4943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4844..4863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5097..5114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5129..5268
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5194..5213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5354..5376
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5370..5561)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatggctctt aacacaccac aaattacgcc gacaaaaaag
      181 ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa
      241 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      301 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      361 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      421 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      481 gatgcgataa cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg
      541 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      601 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      661 gttgcttcta taaaggtaca ggattatgcc catatccagg ctggtgtcgg cttgttctct
      721 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
      781 tgggagcgat tttatccgca gcgccccgct acgcgtaggg acgagaaact tcgtgtctgc
      841 ctagggtgcc attttaccgg ctatccagat cagcgtggtc gctgcccgcg ttgccatatc
      901 ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct gggcggcgct gttagcgtca
      961 atcgttttat tgttacctgc caacctgttg cctatttcta tcatttatct gaacggagga
     1021 cggcaggaag atacaattct ttccggaatt atgtcgctgg caagtagcaa cattgcggtt
     1081 gcaggaatcg tgtttatcgc cagtattctg gtcccgttta ctaaagtgat cgtcatgttc
     1141 actttactgt tgagcattca ttttaaatgc cagcaaggtt tacgcacacg cattctgtta
     1201 ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg acctgtttgt catatcttta
     1261 accatgtcgc tgattaatcg cgatcagatc ctcgctttta ctatgggacc ggctgcgttt
     1321 tatttcggcg cagcggtaat tttgactatt cttgctgtgg aatggctgga cagccgctta
     1381 ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa
     1441 taaacgccgt atctcacctt tctggctgct gccgttcatc gcgctaatga ttgccagttg
     1501 gctgatttgg gacagttatc aggaccgggg taataccgtc accatcgact ttatgtcggc
     1561 ggatggtatt gttccaggcc gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt
     1621 gcaggacatc agcctcagcg acgatcttcg taagattgaa gtcaaggtca gcatcaagtc
     1681 cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg ctggtgacgc caaaagcatc
     1741 gttggcaggt gtctccgggc tggacgccct cgtcggtggg aactatatcg gcatgatgcc
     1801 cgggaaaggt aaagagcagg atcactttgt cgcactcgat acccaaccga aatatcggct
     1861 ggacaatggc gatctgatga tccacctgca agcccccgat ctcggttcgc tgaacagcgg
     1921 ttcattggtc tatttccgca agatcccggt gggaaaagtc tacgactatg ccatcaatcc
     1981 caacaagcaa ggcgtggtga ttgatgtcct gatcgagcgg cgttttaccg atctggtgaa
     2041 aaaaggtagc cgtttctgga acgtttccgg cgttgatgcc aacgtcagta tcagtggcgc
     2101 gaaggtgaaa ctggaaagct tggcggcact ggttaacggt gcgattgcct tcgattcacc
     2161 agaagagtcg aaacctgccg aggcggaaga tacctttggt ctgtatgaag atctggccca
     2221 cagccagcgt ggcgtaataa taaaactgga actgccgagt ggggccggat taaccgccga
     2281 ctcgacgccg ttaatgtatc aggggctgga agtcggacag ctgactaaac tggatttaaa
     2341 tcctggtggt aaagtcacag gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg
     2401 tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc cttagcgatg ctaatctcag
     2461 cgccctgctg accggcaaaa ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga
     2521 attcgttgtt gtgccaggcg aaaaagcact gctgcatgaa cctgatggct taagcattat
     2581 tgttgaagcg ccggaagccg gttcgttagg catcggtacg cctgtgctgt tccgtggtct
     2641 ggaagtcggt acggttacag gaatgacgct ggggacattg tcagatcgcg tgatgattgc
     2701 gatgcgcatc agtaaacgct atcaacacct ggtgcgtaac aattccgtct tctggttggc
     2761 atcgggttac agtctggact ttggtctgac gggcggcgta gtgaaaaccg gcacctttaa
     2821 ccagtttatc cgtggcggca tcgccttcgc cacgcctccg ggtacgccac tggcaccgaa
     2881 agcccaggaa ggcaaacact tcctgttgca ggaaagtgaa ccgaaagagt ggcgtgaatg
     2941 gggaactgcg cttcccaaat aatgcccacc tcgagcagat ccggctgcta acaaagcccg
     3001 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3061 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gataattctt
     3121 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3181 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3241 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3301 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3361 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3421 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3481 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3541 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     3601 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3661 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3721 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3781 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3841 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     3901 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3961 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4021 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4081 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4141 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4201 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4261 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4321 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4381 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4441 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4501 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4561 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4621 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4681 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4741 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4801 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4861 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4921 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4981 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5041 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5101 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     5161 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5221 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     5281 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5341 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5401 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     5461 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     5521 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     5581 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     5641 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     5701 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     5761 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     5821 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     5881 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     5941 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     6001 gacaggag
//
