LOCUS       sequence-334320-        8685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expresson of LetA-[LetB PLL6->4] 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 8685)
  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 8685)
  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: pBEL2252
FEATURES             Location/Qualifiers
     source          1..8685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..879)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        906..1190
                     /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     1113..1132
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1227..1250
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1263..1283
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     primer_bind     complement(5301..5318)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5471..5557
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5649..5676
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5695..5786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5787..5855,5856..6647)
                     /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(6005..6024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6689..7144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6776..6795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6986..7007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      7255..7842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7743..7762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(7939..7972)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     8144..8161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8327..8349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8487..8506
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
       61 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
      121 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
      181 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
      241 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
      301 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
      361 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
      421 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
      481 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
      541 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
      601 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
      661 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
      721 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
      781 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
      841 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
      901 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
      961 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     1021 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     1081 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     1141 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     1201 ttgggctagc cgaagggtga attatgcatc accatcacca ccatcaccat cagcatcaac
     1261 acgaaaatct gtattttcag atggctctta acacaccaca aattacgccg acaaaaaaga
     1321 taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt tgcccgcaat
     1381 gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat tgtccgcgct
     1441 gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg gcaatggcct
     1501 tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat atctggctgt
     1561 taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc aaacagggcg
     1621 atgcgataac ggggtcgatg gtctttttct gcgttattgg tgccccgctc attctggtga
     1681 cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt ccggtactgc
     1741 tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc ggcattggcg
     1801 ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc ttgttctctt
     1861 ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc gaagaactgt
     1921 gggagcgatt ttatccgcag cgccccgcta cgcgtaggga cgagaaactt cgtgtctgtc
     1981 ttgggtgcca ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt tgccatatcc
     2041 cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg ttagcgtcaa
     2101 tcgttttatt gttacctgcc aacctgttgc ctatttctat catttatctg aacggaggac
     2161 ggcaggaaga tacaattctt tccggaatta tgtcgctggc aagtagcaac attgcggttg
     2221 caggaatcgt gtttatcgcc agtattctgg taccgtttac taaagtgatc gtcatgttca
     2281 ctttactgtt gagcattcat tttaaatgcc agcaaggttt acgcacacgc attctgttac
     2341 tgcggatggt gacctggatt ggtcgctggt cgatgctcga cctgtttgtc atatctttaa
     2401 ccatgtcgct gattaatcgc gatcagatcc tcgcttttac tatgggaccg gctgcgtttt
     2461 atttcggcgc agcggtaatt ttgactattc ttgctgtgga atggctggac agccgcttac
     2521 tttgggatgc acatgagtca ggaaacgccc gcttcgacga ctgaagcgca gattaaaaat
     2581 aaacgccgta tctcaccttt ctggctgctg ccgttcatcg cgctaatgat tgccagttgg
     2641 ctgatttggg acagttatca ggaccggggt aataccgtca ccatcgactt tatgtcggcg
     2701 gatggtattg ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt cggaacagtg
     2761 caggatatca gcctcagcga cgatcttcgt aagattgaag tcaaggtcag catcaagtcc
     2821 gatatgaaag atgcgctgcg cgaagagact cagttctggc tggtgacgcc aaaagcatcg
     2881 ttggcaggtg tctccgggct ggacgccctc gtcggtggga actatatcgg catgatgccg
     2941 ggtaaaggta aagagcagga tcactttgtc gcactcgata cccaaccgaa atatcggctg
     3001 gacaatggcg atctgatgat ccacctgcaa gcccccgatc tcggttcgct gaacagcggt
     3061 tcattggtct atttccgcaa gatcccggtg ggaaaagtct acgactatgc catcaatccc
     3121 aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc gttttaccga tctggtgaaa
     3181 aaaggtagcc gtttctggaa cgtttccggc gttgatgcca acgtcagtat cagtggcgcg
     3241 aaggtgaaac tggaaagcct ggcggcactg gttaacggtg cgattgcctt cgattcacca
     3301 gaagagtcga aacctgccga ggcggaagat acctttggtc tgtatgaaga tctggcccac
     3361 agccagcgtg gcgtaataat aaaactggaa ctgccgagtg gggccggatt aaccgccgac
     3421 tcgacgccgt taatgtatca ggggctggaa gtcggacagc tgactaaact ggatttaaat
     3481 cctggtggta aagtcacagg ggaaatgacc gttgatccca gcgtcgttac cctgcttcgt
     3541 gaaaataccc gcatcgaatt acgcaacccg aaattatccc ttagcgatgc taatctcagc
     3601 gccctgctga ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc acgcaaagag
     3661 ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac ctgatgttct gacgctgacc
     3721 ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc cgctcattct tcacggcgtg
     3781 caggtaggcc aggttatcga tcgtaaactc accagcaaag gcgtcacctt taccgtcgcc
     3841 atcgagcctc agcatcgcga actggtaaaa ggcgatagca aatttgtcgt caacagccgt
     3901 gtcgatgtga aggtggggct ggatggcgtt gagtttctcg gtgccagcgc ctcagaatgg
     3961 atcaatggcg ggatacgtat tctgccgggc gataaaggtg agatgaaagc cagctatcca
     4021 ctgtatgcca atttggaaaa agcgctggag aacagcctta gcgatttacc caccacaacc
     4081 gtgagtttga gtgcagagac gctgccggat gtgcaggcag gatcggtagt gctgtaccgt
     4141 aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt tgatatcgat
     4201 ctgcatatta agccggagta tcgcaacctt ctgaccagca atagcgtgtt ctgggcagaa
     4261 ggcggggcga aagttcagct gaatggtagt ggtctgaccg tacaggcatc cccgctctcc
     4321 agagcattaa agggagccat tagcttcgac aacctcagcg gtgccagcgc cagtcagcgt
     4381 aaaggcgata agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt tggcgggcag
     4441 attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat tcgctatctc
     4501 ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa tgaagtacag
     4561 gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg tacgcgcttc
     4621 tcagtggtca gccgtgtcga tgtgaaggtg gggctggatg gcgttgagtt tctcggtgcc
     4681 agcgcctcag aatggatcaa tggcccgtat atcaacgtcg aaccaggccg gggcaatcct
     4741 cgccgcgact ttgaattaca agaggccacc attactgatt cgcgttacct ggatggctta
     4801 agcattattg ttgaagcgcc ggaagccggt tcgttaggca tcggtacgcc tgtgctgttc
     4861 cgtggtctgg aagtcggtac ggttacagga atgacgctgg ggacattgtc agatcgcgtg
     4921 atgattgcga tgcgcatcag taaacgctat caacacctgg tgcgtaacaa ttccgtcttc
     4981 tggttggcat cgggttacag tctggacttt ggtctgacgg gcggcgtagt gaaaaccggc
     5041 acctttaacc agtttatccg tggcggcatc gccttcgcca cgcctccggg tacgccactg
     5101 gcaccgaaag cccaggaagg caaacacttc ctgttgcagg aaagtgaacc gaaagagtgg
     5161 cgtgaatggg gaactgcgct tcccaaataa gtaccaaatt ccagaaaaga gacgctgaaa
     5221 agcgtctttt ttcgttttgg tcctggtctg aggaatggtt gtaagcttgg ctgttttggc
     5281 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     5341 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     5401 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     5461 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     5521 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     5581 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     5641 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt
     5701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6001 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     6061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6301 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     6361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6661 tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg
     6721 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     6781 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     6841 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     6901 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     6961 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     7021 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     7081 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     7141 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7201 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7261 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7321 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gctcagcaga
     7381 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     7441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttagcttg gctggacgta
     7921 aactcctctt cagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg
     7981 ttattgaata tgatcggaat tgggctgcag gaattcgata gcttggctgc aggtcgacct
     8041 cgaggacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     8101 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     8161 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     8221 ccgcataggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     8281 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     8341 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agaaggagat ggcgcccaac
     8401 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     8461 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     8521 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg ctcatgtttg
     8581 acagcttatc atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     8641 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaa
//
