LOCUS       sequence-334285-        8682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expresson of LetA-[LetB PLL6->7] operon from E. coli. N-terminal
            his tag on LetA, araBAD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8682)
  AUTHORS   Vieni C, Coudray N, Isom GL, Bhabha G, Ekiert DC
  TITLE     Role of Ring6 in the Function of the E. coli MCE Protein LetB.
  JOURNAL   J Mol Biol. 2022 Jan 22;434(7):167463. doi: 
            10.1016/j.jmb.2022.167463.
  PUBMED    35077766
REFERENCE   2  (bases 1 to 8682)
  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"; journalName: "J Mol 
            Biol."; date: "2022-01-22"; volume: "434(7):167463. doi"; pages: " 
            10.1016/j.jmb.2022.167463"
COMMENT     Sequence Label: pBEL2253
FEATURES             Location/Qualifiers
     source          1..8682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..61)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      214..300
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      392..419
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        438..529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(530..598,599..1390)
                     /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(748..767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1432..1887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1519..1538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1729..1750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1998..2585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2486..2505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2682..2715)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2887..2904
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3070..3092
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3230..3249
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3429..4307)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4334..4618
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4541..4560
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4655..4678
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4691..4711
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
ORIGIN
        1 gttgtaagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
       61 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
      121 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
      181 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
      241 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      301 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
      361 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
      421 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      481 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      541 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      601 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      661 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      721 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
      781 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      841 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      901 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      961 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1021 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     1081 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1141 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1201 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1261 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1321 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1381 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct
     1441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1501 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1621 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     1681 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1741 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     1801 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1861 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     1921 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1981 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2101 ccgaaggtaa ctggctcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     2161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2641 acatgttagc ttggctggac gtaaactcct cttcagacct aataacttcg tatagcatac
     2701 attatacgaa gttatattaa gggttattga atatgatcgg aattgggctg caggaattcg
     2761 atagcttggc tgcaggtcga cctcgaggac atgttctttc ctgcgttatc ccctgattct
     2821 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2881 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     2941 acgcatctgt gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa
     3001 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3061 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3121 ggcagaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     3181 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     3241 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     3301 cgtagaggat ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg
     3361 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
     3421 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     3481 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     3541 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     3601 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     3661 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     3721 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     3781 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     3841 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     3901 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     3961 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     4021 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     4081 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     4141 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     4201 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     4261 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     4321 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     4381 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     4441 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     4501 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     4561 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     4621 ccgttttttt gggctagccg aagggtgaat tatgcatcac catcaccacc atcaccatca
     4681 gcatcaacac gaaaatctgt attttcagat ggctcttaac acaccacaaa ttacgccgac
     4741 aaaaaagata acagtgaggg caatcggcga ggaactgccg cgtggtgatt accaacgttg
     4801 cccgcaatgt gacatgctgt ttagcctgcc cgagataaat tctcatcaaa gtgcctattg
     4861 tccgcgctgt caggcaaaaa ttcgtgacgg gcgcgactgg tcgctaacgc gcctggcggc
     4921 aatggccttc accatgctgt tgcttatgcc gtttgcctgg ggcgaaccgc tgttgcatat
     4981 ctggctgtta ggcattcgta ttgacgccaa cgttatgcaa ggcatctggc aaatgaccaa
     5041 acagggcgat gcgataacgg ggtcgatggt ctttttctgc gttattggtg ccccgctcat
     5101 tctggtgacc tccatagctt atttatggtt tggtaaccga ctgggaatga atttacgtcc
     5161 ggtactgctg atgcttgagc gacttaaaga gtgggtaatg ctcgatatct acctggtcgg
     5221 cattggcgtt gcttctataa aggtacagga ttatgcccat atccaggctg gtgtcggctt
     5281 gttctctttt gtggcgttgg tgattttaac gacggtgacg ttgtcacatc ttaatgtcga
     5341 agaactgtgg gagcgatttt atccgcagcg ccccgctacg cgtagggacg agaaacttcg
     5401 tgtctgtctt gggtgccatt ttaccggcta tccagatcag cgtggtcgct gcccgcgttg
     5461 ccatatcccg ctacgcctgc gtcgccgtca tagtctgcaa aaatgctggg cggcgctgtt
     5521 agcgtcaatc gttttattgt tacctgccaa cctgttgcct atttctatca tttatctgaa
     5581 cggaggacgg caggaagata caattctttc cggaattatg tcgctggcaa gtagcaacat
     5641 tgcggttgca ggaatcgtgt ttatcgccag tattctggta ccgtttacta aagtgatcgt
     5701 catgttcact ttactgttga gcattcattt taaatgccag caaggtttac gcacacgcat
     5761 tctgttactg cggatggtga cctggattgg tcgctggtcg atgctcgacc tgtttgtcat
     5821 atctttaacc atgtcgctga ttaatcgcga tcagatcctc gcttttacta tgggaccggc
     5881 tgcgttttat ttcggcgcag cggtaatttt gactattctt gctgtggaat ggctggacag
     5941 ccgcttactt tgggatgcac atgagtcagg aaacgcccgc ttcgacgact gaagcgcaga
     6001 ttaaaaataa acgccgtatc tcacctttct ggctgctgcc gttcatcgcg ctaatgattg
     6061 ccagttggct gatttgggac agttatcagg accggggtaa taccgtcacc atcgacttta
     6121 tgtcggcgga tggtattgtt ccaggccgta cgcctgttcg ttatcagggc gttgaagtcg
     6181 gaacagtgca ggatatcagc ctcagcgacg atcttcgtaa gattgaagtc aaggtcagca
     6241 tcaagtccga tatgaaagat gcgctgcgcg aagagactca gttctggctg gtgacgccaa
     6301 aagcatcgtt ggcaggtgtc tccgggctgg acgccctcgt cggtgggaac tatatcggca
     6361 tgatgccggg taaaggtaaa gagcaggatc actttgtcgc actcgatacc caaccgaaat
     6421 atcggctgga caatggcgat ctgatgatcc acctgcaagc ccccgatctc ggttcgctga
     6481 acagcggttc attggtctat ttccgcaaga tcccggtggg aaaagtctac gactatgcca
     6541 tcaatcccaa caagcaaggc gtggtgattg atgtcctgat cgagcggcgt tttaccgatc
     6601 tggtgaaaaa aggtagccgt ttctggaacg tttccggcgt tgatgccaac gtcagtatca
     6661 gtggcgcgaa ggtgaaactg gaaagcctgg cggcactggt taacggtgcg attgccttcg
     6721 attcaccaga agagtcgaaa cctgccgagg cggaagatac ctttggtctg tatgaagatc
     6781 tggcccacag ccagcgtggc gtaataataa aactggaact gccgagtggg gccggattaa
     6841 ccgccgactc gacgccgtta atgtatcagg ggctggaagt cggacagctg actaaactgg
     6901 atttaaatcc tggtggtaaa gtcacagggg aaatgaccgt tgatcccagc gtcgttaccc
     6961 tgcttcgtga aaatacccgc atcgaattac gcaacccgaa attatccctt agcgatgcta
     7021 atctcagcgc cctgctgacc ggcaaaacct tcgagctggt gcccggcgat ggcgagccac
     7081 gcaaagagtt cgttgttgtg ccaggcgaaa aagcactgct gcatgaacct gatgttctga
     7141 cgctgaccct gaccgcgccg gaaagttacg gtattgatgc gggtcagccg ctcattcttc
     7201 acggcgtgca ggtaggccag gttatcgatc gtaaactcac cagcaaaggc gtcaccttta
     7261 ccgtcgccat cgagcctcag catcgcgaac tggtaaaagg cgatagcaaa tttgtcgtca
     7321 acagccgtgt cgatgtgaag gtggggctgg atggcgttga gtttctcggt gccagcgcct
     7381 cagaatggat caatggcggg atacgtattc tgccgggcga taaaggtgag atgaaagcca
     7441 gctatccact gtatgccaat ttggaaaaag cgctggagaa cagccttagc gatttaccca
     7501 ccacaaccgt gagtttgagt gcagagacgc tgccggatgt gcaggcagga tcggtagtgc
     7561 tgtaccgtaa atttgaagtt ggtgaagtga ttaccgtgcg tccgcgagct aacgcgtttg
     7621 atatcgatct gcatattaag ccggagtatc gcaaccttct gaccagcaat agcgtgttct
     7681 gggcagaagg cggggcgaaa gttcagctga atggtagtgg tctgaccgta caggcatccc
     7741 cgctctccag agcattaaag ggagccatta gcttcgacaa cctcagcggt gccagcgcca
     7801 gtcagcgtaa aggcgataag cgtattctgt atgcttccga aacagcggcc cgtgcggttg
     7861 gcgggcagat tacgcttcac gctttcgatg ccggaaaact ggcggtcggg atgccaattc
     7921 gctatctcgg tattgatatc gggcaaatcc agacgctgga tctgattacc gcacgcaatg
     7981 aagtacaggc aaaggcggta ctttacccgg aatatgtcca gacctttgct cgcggtggta
     8041 cgcgcttctc agtggtctcg ggttacagtc tggactttgg tctgacgggc ggcgtagtga
     8101 aaaccggcac ctttaaccag tttatccgtg gctatatcaa cgtcgaacca ggccggggca
     8161 atcctcgccg cgactttgaa ttacaagagg ccaccattac tgattcgcgt tacctggatg
     8221 gcttaagcat tattgttgaa gcgccggaag ccggttcgtt aggcatcggt acgcctgtgc
     8281 tgttccgtgg tctggaagtc ggtacggtta caggaatgac gctggggaca ttgtcagatc
     8341 gcgtgatgat tgcgatgcgc atcagtaaac gctatcaaca cctggtgcgt aacaattccg
     8401 tcttctggtt ggcatcgggt tacagtctgg actttggtct gacgggcggc gtagtgaaaa
     8461 ccggcacctt taaccagttt atccgtggcg gcatcgcctt cgccacgcct ccgggtacgc
     8521 cactggcacc gaaagcccag gaaggcaaac acttcctgtt gcaggaaagt gaaccgaaag
     8581 agtggcgtga atggggaact gcgcttccca aataagtacc aaattccaga aaagagacgc
     8641 tgaaaagcgt cttttttcgt tttggtcctg gtctgaggaa tg
//
