LOCUS       sequence-345244-        8664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expresson of LetA-[LetB PLL3->6] 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 8664)
  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 8664)
  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: pBEL2265
FEATURES             Location/Qualifiers
     source          1..8664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..294
                     /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     217..236
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..354
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             367..387
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     primer_bind     complement(4384..4401)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4554..4640
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4732..4759
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4778..4869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4870..4938,4939..5730)
                     /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(5088..5107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5772..6227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5859..5878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6069..6090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6338..6925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6826..6845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(7022..7055)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     7227..7244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7410..7432
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7570..7589
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7769..8647)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
       61 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
      121 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
      181 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
      241 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
      301 ttttttgggc tagccgaagg gtgaattatg catcaccatc accaccatca ccatcagcat
      361 caacacgaaa atctgtattt tcagatggct cttaacacac cacaaattac gccgacaaaa
      421 aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca acgttgcccg
      481 caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc ctattgtccg
      541 cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct ggcggcaatg
      601 gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt gcatatctgg
      661 ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat gaccaaacag
      721 ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc gctcattctg
      781 gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt acgtccggta
      841 ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct ggtcggcatt
      901 ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt cggcttgttc
      961 tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa tgtcgaagaa
     1021 ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa acttcgtgtc
     1081 tgtcttgggt gccattttac cggctatcca gatcagcgtg gtcgctgccc gcgttgccat
     1141 atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc gctgttagcg
     1201 tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta tctgaacgga
     1261 ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag caacattgcg
     1321 gttgcaggaa tcgtgtttat cgccagtatt ctggtaccgt ttactaaagt gatcgtcatg
     1381 ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac acgcattctg
     1441 ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt tgtcatatct
     1501 ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg accggctgcg
     1561 ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct ggacagccgc
     1621 ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag cgcagattaa
     1681 aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa tgattgccag
     1741 ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg actttatgtc
     1801 ggcggatggt attgttccag gccgtacgcc tgttcgttat cagggcgttg aagtcggaac
     1861 agtgcaggat atcagcctca gcgacgatct tcgtaagatt gaagtcaagg tcagcatcaa
     1921 gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga cgccaaaagc
     1981 atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata tcggcatgat
     2041 gccgggtaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac cgaaatatcg
     2101 gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt cgctgaacag
     2161 cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact atgccatcaa
     2221 tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta ccgatctggt
     2281 gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca gtatcagtgg
     2341 cgcgaaggtg aaactggaaa gcctggcggc actggttaac ggtgcgattg ccttcgattc
     2401 accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg aagatctggc
     2461 ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg gattaaccgc
     2521 cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta aactggattt
     2581 aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg ttaccctgct
     2641 tcgtgaaaat acccgcatcg aattacgcaa cccgcaaatt tcggcagctg gcgttgagca
     2701 tcttgatact atcctccagg gcaaaacctt cgagctggtg cccggcgatg gcgagccacg
     2761 caaagagttc gttgttgtgc caggcgaaaa agcactgctg catgaacctg atgttctgac
     2821 gctgaccctg accgcgccgg aaagttacgg tattgatgcg ggtcagccgc tcattcttca
     2881 cggcgtgcag gtaggccagg ttatcgatcg taaactcacc agcaaaggcg tcacctttac
     2941 cgtcgccatc gagcctcagc atcgcgaact ggtaaaaggc gatagcaaat ttgtcgtcaa
     3001 cagccgtgtc gatgtgaagg tggggctgga tggcgttgag tttctcggtg ccagcgcctc
     3061 agaatggatc aatggcggga tacgtattct gccgggcgat aaaggtgaga tgaaagccag
     3121 ctatccactg tatgccaatt tggaaaaagc gctggagaac agccttagcg atttacccac
     3181 cacaaccgtg agtttgagtg cagagacgct gccggatgtg caggcaggat cggtagtgct
     3241 gtaccgtaaa tttgaagttg gtgaagtgat taccgtgcgt ccgcgagcta acgcgtttga
     3301 tatcgatctg catattaagc cggagtatcg caaccttctg accagcaata gcgtgttctg
     3361 ggcagaaggc ggggcgaaag ttcagctgaa tggtagtggt ctgaccgtac aggcatcccc
     3421 gctctccaga gcattaaagg gagccattag cttcgacaac ctcagcggtg ccagcgccag
     3481 tcagcgtaaa ggcgataagc gtattctgta tgcttccgaa acagcggccc gtgcggttgg
     3541 cgggcagatt acgcttcacg ctttcgatgc cggaaaactg gcggtcggga tgccaattcg
     3601 ctatctcggt attgatatcg ggcaaatcca gacgctggat ctgattaccg cacgcaatga
     3661 agtacaggca aaggcggtac tttacccgga atatgtccag acctttgctc gcggtggtac
     3721 gcgcttctca gtggtcacac cgcaaatttc ggcagctggc gttgagcatc ttgatactat
     3781 cctccagccg tatatcaacg tcgaaccagg ccggggcaat cctcgccgcg actttgaatt
     3841 acaagaggcc accattactg attcgcgtta cctggatggc ttaagcatta ttgttgaagc
     3901 gccggaagcc ggttcgttag gcatcggtac gcctgtgctg ttccgtggtc tggaagtcgg
     3961 tacggttaca ggaatgacgc tggggacatt gtcagatcgc gtgatgattg cgatgcgcat
     4021 cagtaaacgc tatcaacacc tggtgcgtaa caattccgtc ttctggttgg catcgggtta
     4081 cagtctggac tttggtctga cgggcggcgt agtgaaaacc ggcaccttta accagtttat
     4141 ccgtggcggc atcgccttcg ccacgcctcc gggtacgcca ctggcaccga aagcccagga
     4201 aggcaaacac ttcctgttgc aggaaagtga accgaaagag tggcgtgaat ggggaactgc
     4261 gcttcccaaa taagtaccaa attccagaaa agagacgctg aaaagcgtct tttttcgttt
     4321 tggtcctggt ctgaggaatg gttgtaagct tggctgtttt ggcggatgag agaagatttt
     4381 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     4441 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     4501 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     4561 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     4621 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     4681 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     4741 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     4801 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4861 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4921 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4981 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5041 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5101 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5161 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5221 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5281 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5341 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5401 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5461 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5521 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5581 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5641 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5701 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5761 tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5821 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     6001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     6121 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6181 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa aggatctagg
     6241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6301 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6421 aagagctacc aactcttttt ccgaaggtaa ctggctcagc agagcgcaga taccaaatac
     6481 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6961 cttttgctgg ccttttgctc acatgttagc ttggctggac gtaaactcct cttcagacct
     7021 aataacttcg tatagcatac attatacgaa gttatattaa gggttattga atatgatcgg
     7081 aattgggctg caggaattcg atagcttggc tgcaggtcga cctcgaggac atgttctttc
     7141 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7201 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7261 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtcatggc
     7321 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     7381 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     7441 gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc cggccacggg
     7501 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     7561 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     7621 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
     7681 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
     7741 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
     7801 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     7861 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     7921 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     7981 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     8041 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     8101 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     8161 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     8221 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     8281 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     8341 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     8401 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     8461 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     8521 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     8581 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     8641 cagccatact tttcatactc ccgc
//
