LOCUS       sequence-309654-        5651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 3 ring (rings 167) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5651)
  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 5651)
  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: pBEL2006
FEATURES             Location/Qualifiers
     source          1..5651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     229..248
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        229..247
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             279..301
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2837..2855)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2851..2898
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2943..2961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3029..3133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3134..3202,3203..3994)
                     /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(3352..3371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4165..4753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4654..4673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4907..4924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4939..5078
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5004..5023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5164..5186
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5180..5371)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
       61 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
      121 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
      181 acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaatta atacgactca
      241 ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
      301 atatacatat ggctcttaac acaccacaaa ttacgccgac aaaaaagata acagtgaggg
      361 caatcggcga ggaactgccg cgtggtgatt accaacgttg cccgcaatgt gacatgctgt
      421 ttagcctgcc cgagataaat tctcatcaaa gtgcctattg tccgcgctgt caggcaaaaa
      481 ttcgtgacgg gcgcgactgg tcgctaacgc gcctggcggc aatggccttc accatgctgt
      541 tgcttatgcc gtttgcctgg ggcgaaccgc tgttgcatat ctggctgtta ggcattcgta
      601 ttgacgccaa cgttatgcaa ggcatctggc aaatgaccaa acagggcgat gcgataacgg
      661 ggtcgatggt ctttttctgc gttattggtg ccccgctcat tctggtgacc tccatagctt
      721 atttatggtt tggtaaccga ctgggaatga atttacgtcc ggtactgctg atgcttgagc
      781 gacttaaaga gtgggtaatg ctcgatatct acctggtcgg cattggcgtt gcttctataa
      841 aggtacagga ttatgcccat atccaggctg gtgtcggctt gttctctttt gtggcgttgg
      901 tgattttaac gacggtgacg ttgtcacatc ttaatgtcga agaactgtgg gagcgatttt
      961 atccgcagcg ccccgctacg cgtagggacg agaaacttcg tgtctgccta gggtgccatt
     1021 ttaccggcta tccagatcag cgtggtcgct gcccgcgttg ccatatcccg ctacgcctgc
     1081 gtcgccgtca tagtctgcaa aaatgctggg cggcgctgtt agcgtcaatc gttttattgt
     1141 tacctgccaa cctgttgcct atttctatca tttatctgaa cggaggacgg caggaagata
     1201 caattctttc cggaattatg tcgctggcaa gtagcaacat tgcggttgca ggaatcgtgt
     1261 ttatcgccag tattctggtc ccgtttacta aagtgatcgt catgttcact ttactgttga
     1321 gcattcattt taaatgccag caaggtttac gcacacgcat tctgttactg cggatggtga
     1381 cctggattgg tcgctggtcg atgctcgacc tgtttgtcat atctttaacc atgtcgctga
     1441 ttaatcgcga tcagatcctc gcttttacta tgggaccggc tgcgttttat ttcggcgcag
     1501 cggtaatttt gactattctt gctgtggaat ggctggacag ccgcttactt tgggatgcac
     1561 atgagtcagg aaacgcccgc ttcgacgact gaagcgcaga ttaaaaataa acgccgtatc
     1621 tcacctttct ggctgctgcc gttcatcgcg ctaatgattg ccagttggct gatttgggac
     1681 agttatcagg accggggtaa taccgtcacc atcgacttta tgtcggcgga tggtattgtt
     1741 ccaggccgta cgcctgttcg ttatcagggc gttgaagtcg gaacagtgca ggacatcagc
     1801 ctcagcgacg atcttcgtaa gattgaagtc aaggtcagca tcaagtccga tatgaaagat
     1861 gcgctgcgcg aagagactca gttctggctg gtgacgccaa aagcatcgtt ggcaggtgtc
     1921 tccgggctgg acgccctcgt cggtgggaac tatatcggca tgatgcccgg gaaaggtaaa
     1981 gagcaggatc actttgtcgc actcgatacc caaccgaaat atcggctgga caatggcgat
     2041 gggcagatta cgcttcacgc tttcgatgcc ggaaaactgg cggtcgggat gccaattcgc
     2101 tatctcggta ttgacatcgg gcaaatccag acgctggatc tgattaccgc acgcaatgaa
     2161 gtacaggcaa aggcggtact ttacccggaa tatgtccaga cctttgctcg cggtggtacg
     2221 cgcttctcag tggtcacacc gcaaatttcg gcagctggcg ttgagcatct tgatactatc
     2281 ctccagccgt atatcaacgt cgaaccaggc cggggcaatc ctcgccgcga ctttgaatta
     2341 caagaggcca ccattactga ttcgcggtac ctggatggct taagcattat tgttgaagcg
     2401 ccggaagccg gttcgttagg catcggtacg cctgtgctgt tccgtggtct ggaagtcggt
     2461 acggttacag gaatgacgct ggggacattg tcagatcgcg tgatgattgc gatgcgcatc
     2521 agtaaacgct atcaacacct ggtgcgtaac aattccgtct tctggttggc atcgggttac
     2581 agtctggact ttggtctgac gggcggcgta gtgaaaaccg gcacctttaa ccagtttatc
     2641 cgtggcggca tcgccttcgc cacgcctccg ggtacgccac tggcaccgaa agcccaggaa
     2701 ggcaaacact tcctgttgca ggaaagtgaa ccgaaagagt ggcgtgaatg gggaactgcg
     2761 cttcccaaat aatgcccacc tcgagcagat ccggctgcta acaaagcccg aaaggaagct
     2821 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2881 gtcttgaggg gttttttgct gaaaggagga actatatccg gataattctt gaagacgaaa
     2941 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3001 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3061 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3121 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3181 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3241 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3301 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3361 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     3421 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3481 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3541 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3601 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3661 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3721 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3781 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3841 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3901 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3961 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4021 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     4321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4861 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4921 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4981 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     5041 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     5101 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5161 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     5221 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     5281 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     5341 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     5401 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     5461 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     5521 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     5581 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     5641 ctgacttccg c
//
