LOCUS       sequence-310583-        7043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB PLL4 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  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 7043)
  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: pBEL1889
FEATURES             Location/Qualifiers
     source          1..7043
                     /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(4222..4240)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4236..4283
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4328..4346)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4414..4518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4519..4587,4588..5379)
                     /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(4737..4756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5550..6138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6039..6058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6292..6309
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6324..6463
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6389..6408)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6549..6571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6565..6756)
                     /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 cctgatgttc tgacgctgac cctgaccgcg ccggaaagtt
     2761 acggtattga tgcgggtcag ccgctcattc ttcacggcgt gcaggtaggc caggttatcg
     2821 atcgtaaact caccagcaaa ggcgtcacct ttaccgtcgc catcgagcct cagcatcgcg
     2881 aactggtaaa aggcgatagc aaatttgtcg tcaacagccg tgtcgatctc ggtgccagcg
     2941 cctcagaatg gatcaatggc gggatccgta ttctgccggg cgataaaggt gagatgaaag
     3001 ccagctatcc actgtatgcc aatttggaaa aagcgctgga gaacagcctt agcgatttac
     3061 ccaccacaac cgtgagtttg agtgcagaga cgctgccgga tgtgcaggca ggatcggtag
     3121 tgctgtaccg taaatttgaa gttggtgaag tgattaccgt gcgtccgcga gctaacgcgt
     3181 ttgacatcga tctgcatatt aagccggagt atcgcaacct tctgaccagc aatagcgtgt
     3241 tctgggcaga aggcggggcg aaagttcagc tgaatggtag tggtctgacc gtacaggcat
     3301 ccccgctctc cagagctcta aagggagcca ttagcttcga caacctcagc ggtgccagcg
     3361 ccagtcagcg taaaggcgat aagcgtattc tgtatgcttc cgaaacagcg gcccgtgcgg
     3421 ttggcgggca gattacgctt cacgctttcg atgccggaaa actggcggtc gggatgccaa
     3481 ttcgctatct cggtattgac atcgggcaaa tccagacgct ggatctgatt accgcacgca
     3541 atgaagtaca ggcaaaggcg gtactttacc cggaatatgt ccagaccttt gctcgcggtg
     3601 gtacgcgctt ctcagtggtc acaccgcaaa tttcggcagc tggcgttgag catcttgata
     3661 ctatcctcca gccgtatatc aacgtcgaac caggccgggg caatcctcgc cgcgactttg
     3721 aattacaaga ggccaccatt actgattcgc ggtacctgga tggcttaagc attattgttg
     3781 aagcgccgga agccggttcg ttaggcatcg gtacgcctgt gctgttccgt ggtctggaag
     3841 tcggtacggt tacaggaatg acgctgggga cattgtcaga tcgcgtgatg attgcgatgc
     3901 gcatcagtaa acgctatcaa cacctggtgc gtaacaattc cgtcttctgg ttggcatcgg
     3961 gttacagtct ggactttggt ctgacgggcg gcgtagtgaa aaccggcacc tttaaccagt
     4021 ttatccgtgg cggcatcgcc ttcgccacgc ctccgggtac gccactggca ccgaaagccc
     4081 aggaaggcaa acacttcctg ttgcaggaaa gtgaaccgaa agagtggcgt gaatggggaa
     4141 ctgcgcttcc caaataatgc ccacctcgag cagatccggc tgctaacaaa gcccgaaagg
     4201 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     4261 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa ttcttgaaga
     4321 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4381 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4441 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4501 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4621 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4681 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4741 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     4801 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4921 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5041 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     5101 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5341 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5401 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6241 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6301 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6361 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6421 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     6481 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6541 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6601 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     6661 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     6721 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     6781 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     6841 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     6901 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     6961 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     7021 gggcgctgac ttccgcgttt cca
//
