LOCUS       sequence-309656-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with 4 ring (rings 1567) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  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 6011)
  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: pBEL2007
FEATURES             Location/Qualifiers
     source          1..6011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     147..166
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        147..165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             197..219
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3115..3133)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3129..3176
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3221..3239)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3307..3411
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3412..3480,3481..4272)
                     /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(3630..3649)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4443..5031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4932..4951
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5185..5202
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5217..5356
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5282..5301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5442..5464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5458..5649)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
       61 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
      121 tgcccgagat ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
      181 tccctctaga aataattttg tttaacttta agaaggagat atacatatgg ctcttaacac
      241 accacaaatt acgccgacaa aaaagataac agtgagggca atcggcgagg aactgccgcg
      301 tggtgattac caacgttgcc cgcaatgtga catgctgttt agcctgcccg agataaattc
      361 tcatcaaagt gcctattgtc cgcgctgtca ggcaaaaatt cgtgacgggc gcgactggtc
      421 gctaacgcgc ctggcggcaa tggccttcac catgctgttg cttatgccgt ttgcctgggg
      481 cgaaccgctg ttgcatatct ggctgttagg cattcgtatt gacgccaacg ttatgcaagg
      541 catctggcaa atgaccaaac agggcgatgc gataacgggg tcgatggtct ttttctgcgt
      601 tattggtgcc ccgctcattc tggtgacctc catagcttat ttatggtttg gtaaccgact
      661 gggaatgaat ttacgtccgg tactgctgat gcttgagcga cttaaagagt gggtaatgct
      721 cgatatctac ctggtcggca ttggcgttgc ttctataaag gtacaggatt atgcccatat
      781 ccaggctggt gtcggcttgt tctcttttgt ggcgttggtg attttaacga cggtgacgtt
      841 gtcacatctt aatgtcgaag aactgtggga gcgattttat ccgcagcgcc ccgctacgcg
      901 tagggacgag aaacttcgtg tctgcctagg gtgccatttt accggctatc cagatcagcg
      961 tggtcgctgc ccgcgttgcc atatcccgct acgcctgcgt cgccgtcata gtctgcaaaa
     1021 atgctgggcg gcgctgttag cgtcaatcgt tttattgtta cctgccaacc tgttgcctat
     1081 ttctatcatt tatctgaacg gaggacggca ggaagataca attctttccg gaattatgtc
     1141 gctggcaagt agcaacattg cggttgcagg aatcgtgttt atcgccagta ttctggtccc
     1201 gtttactaaa gtgatcgtca tgttcacttt actgttgagc attcatttta aatgccagca
     1261 aggtttacgc acacgcattc tgttactgcg gatggtgacc tggattggtc gctggtcgat
     1321 gctcgacctg tttgtcatat ctttaaccat gtcgctgatt aatcgcgatc agatcctcgc
     1381 ttttactatg ggaccggctg cgttttattt cggcgcagcg gtaattttga ctattcttgc
     1441 tgtggaatgg ctggacagcc gcttactttg ggatgcacat gagtcaggaa acgcccgctt
     1501 cgacgactga agcgcagatt aaaaataaac gccgtatctc acctttctgg ctgctgccgt
     1561 tcatcgcgct aatgattgcc agttggctga tttgggacag ttatcaggac cggggtaata
     1621 ccgtcaccat cgactttatg tcggcggatg gtattgttcc aggccgtacg cctgttcgtt
     1681 atcagggcgt tgaagtcgga acagtgcagg acatcagcct cagcgacgat cttcgtaaga
     1741 ttgaagtcaa ggtcagcatc aagtccgata tgaaagatgc gctgcgcgaa gagactcagt
     1801 tctggctggt gacgccaaaa gcatcgttgg caggtgtctc cgggctggac gccctcgtcg
     1861 gtgggaacta tatcggcatg atgccgggta aaggtaaaga gcaggatcac tttgtcgcac
     1921 tcgataccca accgaaatat cggctggaca atggcgatac aaccgtgagt ttgagtgcag
     1981 agacgctgcc ggatgtgcag gcaggatcgg tagtgctgta ccgtaaattt gaagttggtg
     2041 aagtgattac cgtgcgtccg cgagctaacg cgtttgatat cgatctgcat attaagccgg
     2101 agtatcgcaa ccttctgacc agcaatagcg tgttctgggc agaaggcggg gcgaaagttc
     2161 agctgaatgg tagtggtctg accgtacagg catccccgct ctccagagca ttaaagggag
     2221 ccattagctt cgacaacctc agcggtgcca gcgccagtca gcgtaaaggc gataagcgta
     2281 ttctgtatgc ttccgaaaca gcggcccgtg cggttggcgg gcagattacg cttcacgctt
     2341 tcgatgccgg aaaactggcg gtcgggatgc caattcgcta tctcggtatt gatatcgggc
     2401 aaatccagac gctggatctg attaccgcac gcaatgaagt acaggcaaag gcggtacttt
     2461 acccggaata tgtccagacc tttgctcgcg gtggtacgcg cttctcagtg gtcacaccgc
     2521 aaatttcggc agctggcgtt gagcatcttg atactatcct ccagccgtat atcaacgtcg
     2581 aaccaggccg gggcaatcct cgccgcgact ttgaattaca agaggccacc attactgatt
     2641 cgcgttacct ggatggctta agcattattg ttgaagcgcc ggaagccggt tcgttaggca
     2701 tcggtacgcc tgtgctgttc cgtggtctgg aagtcggtac ggttacagga atgacgctgg
     2761 ggacattgtc agatcgcgtg atgattgcga tgcgcatcag taaacgctat caacacctgg
     2821 tgcgtaacaa ttccgtcttc tggttggcat cgggttacag tctggacttt ggtctgacgg
     2881 gcggcgtagt gaaaaccggc acctttaacc agtttatccg tggcggcatc gccttcgcca
     2941 cgcctccggg tacgccactg gcaccgaaag cccaggaagg caaacacttc ctgttgcagg
     3001 aaagtgaacc gaaagagtgg cgtgaatggg gaactgcgct tcccaaataa tgcccacctc
     3061 gagcagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     3121 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     3181 aaggaggaac tatatccgga taattcttga agacgaaagg gcctcgtgat acgcctattt
     3241 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3301 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3361 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3421 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3481 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3541 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3601 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     3661 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3721 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3781 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3841 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3901 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     3961 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4021 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4081 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4141 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4201 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4261 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4321 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4381 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4441 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4501 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4561 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4621 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4681 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4741 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4801 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4861 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4921 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4981 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5041 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5101 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5161 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     5221 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     5281 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     5341 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5401 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5461 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5521 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5581 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5641 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     5701 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     5761 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     5821 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     5881 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     5941 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     6001 gcagcagtcg c
//
