LOCUS       sequence-345259-        8322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of LetA-[Delta-Ring6 LetB] 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 8322)
  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 8322)
  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: pBEL2114
FEATURES             Location/Qualifiers
     source          1..8322
                     /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(4042..4059)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4212..4298
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4390..4417
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4436..4527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4528..4596,4597..5388)
                     /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(4746..4765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5430..5885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5517..5536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5727..5748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5996..6583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6484..6503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(6680..6713)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     6885..6902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7068..7090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7228..7247
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7427..8305)
                     /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 cccgaaatta tcccttagcg atgctaatct
     2701 cagcgccctg ctgaccggca aaaccttcga gctggtgccc ggcgatggcg agccacgcaa
     2761 agagttcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg ttctgacgct
     2821 gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca ttcttcacgg
     2881 cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca cctttaccgt
     2941 cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg tcgtcaacag
     3001 ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca gcgcctcaga
     3061 atggatcaat ggcgggatac gtattctgcc gggcgataaa ggtgagatga aagccagcta
     3121 tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt tacccaccac
     3181 aaccgtgagt ttgagtgcag agacgctgcc ggatgtgcag gcaggatcgg tagtgctgta
     3241 ccgtaaattt gaagttggtg aagtgattac cgtgcgtccg cgagctaacg cgtttgatat
     3301 cgatctgcat attaagccgg agtatcgcaa ccttctgacc agcaatagcg tgttctgggc
     3361 agaaggcggg gcgaaagttc agctgaatgg tagtggtctg accgtacagg catccccgct
     3421 ctccagagca ttaaagggag ccattagctt cgacaacctc agcggtgcca gcgccagtca
     3481 gcgtaaaggc gataagcgta ttctgtatgc ttccgaaaca gcggcccgtg cggttggctt
     3541 aagcattatt gttgaagcgc cggaagccgg ttcgttaggc atcggtacgc ctgtgctgtt
     3601 ccgtggtctg gaagtcggta cggttacagg aatgacgctg gggacattgt cagatcgcgt
     3661 gatgattgcg atgcgcatca gtaaacgcta tcaacacctg gtgcgtaaca attccgtgtt
     3721 ctggttggca tcgggttaca gtctggactt tggtctgacg ggcggcgtag tgaaaaccgg
     3781 cacctttaac cagtttatcc gtggcggcat cgccttcgcc acgcctccgg gtacgccact
     3841 ggcaccgaaa gcccaggaag gcaaacactt cctgttgcag gaaagtgaac cgaaagagtg
     3901 gcgtgaatgg ggaactgcgc ttcccaaata agtaccaaat tccagaaaag agacgctgaa
     3961 aagcgtcttt tttcgttttg gtcctggtct gaggaatggt tgtaagcttg gctgttttgg
     4021 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     4081 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     4141 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     4201 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4261 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     4321 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     4381 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     4441 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4501 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4561 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4621 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4681 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4741 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     4801 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4861 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4921 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4981 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5041 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5101 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5161 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5221 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5281 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5341 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5401 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     5461 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5521 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5581 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5641 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5701 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5761 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5821 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     5881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggctcagcag
     6121 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6361 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttagctt ggctggacgt
     6661 aaactcctct tcagacctaa taacttcgta tagcatacat tatacgaagt tatattaagg
     6721 gttattgaat atgatcggaa ttgggctgca ggaattcgat agcttggctg caggtcgacc
     6781 tcgaggacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6841 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6901 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6961 accgcatagg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     7021 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     7081 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa
     7141 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     7201 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     7261 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt
     7321 gacagcttat catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa
     7381 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga
     7441 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg
     7501 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga
     7561 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt
     7621 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg
     7681 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt
     7741 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact
     7801 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct
     7861 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat
     7921 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt
     7981 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc
     8041 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac
     8101 ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga
     8161 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt
     8221 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca
     8281 ggggatcatt ttgcgcttca gccatacttt tcatactccc gc
//
