LOCUS       sequence-309828-        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     222..241
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        222..240
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             272..294
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3187..3205)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3201..3248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3293..3311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3379..3483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3484..3552,3553..4344)
                     /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(3702..3721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4515..5103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5004..5023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5257..5274
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5289..5428
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5354..5373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5514..5536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5530..5721)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
       61 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
      121 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
      181 gccaggaccc aacgctgccc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
      241 gagaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
      301 tatggctctt aacacaccac aaattacgcc gacaaaaaag ataacagtga gggcaatcgg
      361 cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa tgtgacatgc tgtttagcct
      421 gcccgagata aattctcatc aaagtgccta ttgtccgcgc tgtcaggcaa aaattcgtga
      481 cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc ttcaccatgc tgttgcttat
      541 gccgtttgcc tggggcgaac cgctgttgca tatctggctg ttaggcattc gtattgacgc
      601 caacgttatg caaggcatct ggcaaatgac caaacagggc gatgcgataa cggggtcgat
      661 ggtctttttc tgcgttattg gtgccccgct cattctggtg acctccatag cttatttatg
      721 gtttggtaac cgactgggaa tgaatttacg tccggtactg ctgatgcttg agcgacttaa
      781 agagtgggta atgctcgata tctacctggt cggcattggc gttgcttcta taaaggtaca
      841 ggattatgcc catatccagg ctggtgtcgg cttgttctct tttgtggcgt tggtgatttt
      901 aacgacggtg acgttgtcac atcttaatgt cgaagaactg tgggagcgat tttatccgca
      961 gcgccccgct acgcgtaggg acgagaaact tcgtgtctgc ctagggtgcc attttaccgg
     1021 ctatccagat cagcgtggtc gctgcccgcg ttgccatatc ccgctacgcc tgcgtcgccg
     1081 tcatagtctg caaaaatgct gggcggcgct gttagcgtca atcgttttat tgttacctgc
     1141 caacctgttg cctatttcta tcatttatct gaacggagga cggcaggaag atacaattct
     1201 ttccggaatt atgtcgctgg caagtagcaa cattgcggtt gcaggaatcg tgtttatcgc
     1261 cagtattctg gtcccgttta ctaaagtgat cgtcatgttc actttactgt tgagcattca
     1321 ttttaaatgc cagcaaggtt tacgcacacg cattctgtta ctgcggatgg tgacctggat
     1381 tggtcgctgg tcgatgctcg acctgtttgt catatcttta accatgtcgc tgattaatcg
     1441 cgatcagatc ctcgctttta ctatgggacc ggctgcgttt tatttcggcg cagcggtaat
     1501 tttgactatt cttgctgtgg aatggctgga cagccgctta ctttgggatg cacatgagtc
     1561 aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa taaacgccgt atctcacctt
     1621 tctggctgct gccgttcatc gcgctaatga ttgccagttg gctgatttgg gacagttatc
     1681 aggaccgggg taataccgtc accatcgact ttatgtcggc ggatggtatt gttccaggcc
     1741 gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt gcaggacatc agcctcagcg
     1801 acgatcttcg taagattgaa gtcaaggtca gcatcaagtc cgatatgaaa gatgcgctgc
     1861 gcgaagagac tcagttctgg ctggtgacgc caaaagcatc gttggcaggt gtctccgggc
     1921 tggacgccct cgtcggtggg aactatatcg gcatgatgcc cgggaaaggt aaagagcagg
     1981 atcactttgt cgcactcgat acccaaccga aatatcggct ggacaatggc gatctgatga
     2041 tccacctgca agcccccgat ctcggttcgc tgaacagcgg ttcattggtc tatttccgca
     2101 agatcccggt gggaaaagtc tacgactatg ccatcaatcc caacaagcaa ggcgtggtga
     2161 ttgatgtcct gatcgagcgg cgttttaccg atctggtgaa aaaaggtagc cgtttctgga
     2221 acgtttccgg cgttgatgcc aacgtcagta tcagtggcgc gaaggtgaaa ctggaaagct
     2281 tggcggcact ggttaacggt gcgattgcct tcgattcacc agaagagtcg aaacctgccg
     2341 aggcggaaga tacctttggt ctgtatgaag atctggccca cagccagcgt ggcgtaataa
     2401 taaaactgga actgccgagt ggggccggat taaccgccga ctcgacgccg ttaatgtatc
     2461 aggggctgga agtcggacag ctgactaaac tggatttaaa tcctggtggt aaagtcacag
     2521 gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg tgaaaatacc cgcatcgaat
     2581 tacgcaaccc gaaattatcc cttagcgatg ctaatctcag cgccctgctg accggcaaaa
     2641 ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga attcgttgtt gtgccaggcg
     2701 aaaaagcact gctgcatgaa cctgatggct taagcattat tgttgaagcg ccggaagccg
     2761 gttcgttagg catcggtacg cctgtgctgt tccgtggtct ggaagtcggt acggttacag
     2821 gaatgacgct ggggacattg tcagatcgcg tgatgattgc gatgcgcatc agtaaacgct
     2881 atcaacacct ggtgcgtaac aattccgtct tctggttggc atcgggttac agtctggact
     2941 ttggtctgac gggcggcgta gtgaaaaccg gcacctttaa ccagtttatc cgtggcggca
     3001 tcgccttcgc cacgcctccg ggtacgccac tggcaccgaa agcccaggaa ggcaaacact
     3061 tcctgttgca ggaaagtgaa ccgaaagagt ggcgtgaatg gggaactgcg cttcccaaat
     3121 aatgcccacc tcgagcagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3181 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     3241 gttttttgct gaaaggagga actatatccg gataattctt gaagacgaaa gggcctcgtg
     3301 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3361 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3421 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3481 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3541 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3601 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3661 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3721 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3781 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3841 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3901 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3961 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4021 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4081 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4141 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4201 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4261 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4321 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4381 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4441 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4501 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4561 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4621 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4681 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4741 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4801 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4861 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4921 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4981 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5041 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5101 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5161 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5221 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5281 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     5341 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     5401 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     5461 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5521 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5581 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5641 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5701 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5761 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5821 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     5881 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     5941 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     6001 cgtttcca
//
