LOCUS       sequence-334331-        8682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expresson of LetA-[LetB PLL6->2] 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: pBEL2251
FEATURES             Location/Qualifiers
     source          1..8682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..74)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      227..313
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      405..432
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        451..542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(543..611,612..1403)
                     /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(761..780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1445..1900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1532..1551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1742..1763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2011..2598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2499..2518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2695..2728)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2900..2917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3083..3105
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3243..3262
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3442..4320)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4347..4631
                     /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     4554..4573
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4668..4691
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4704..4724
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
ORIGIN
        1 ggtctgagga atggttgtaa gcttggctgt tttggcggat gagagaagat tttcagcctg
       61 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
      121 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
      181 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
      241 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      301 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
      361 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
      421 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
      481 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      541 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      601 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      661 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      721 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      781 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      841 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      901 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      961 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1021 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1081 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1141 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1201 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1261 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1321 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1381 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1441 atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1501 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     1681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1741 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     1801 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1861 tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat
     1921 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1981 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2101 accaactctt tttccgaagg taactggctc agcagagcgc agataccaaa tactgtcctt
     2161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2641 tggccttttg ctcacatgtt agcttggctg gacgtaaact cctcttcaga cctaataact
     2701 tcgtatagca tacattatac gaagttatat taagggttat tgaatatgat cggaattggg
     2761 ctgcaggaat tcgatagctt ggctgcaggt cgacctcgag gacatgttct ttcctgcgtt
     2821 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2881 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc
     3001 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3061 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3121 ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     3181 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     3241 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     3301 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg
     3361 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     3421 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     3481 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     3541 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     3601 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     3661 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     3721 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     3781 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     3841 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     3901 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     3961 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     4021 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     4081 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     4141 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     4201 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     4261 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     4321 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
     4381 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
     4441 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
     4501 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
     4561 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac
     4621 tgtttctcca tacccgtttt tttgggctag ccgaagggtg aattatgcat caccatcacc
     4681 accatcacca tcagcatcaa cacgaaaatc tgtattttca gatggctctt aacacaccac
     4741 aaattacgcc gacaaaaaag ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg
     4801 attaccaacg ttgcccgcaa tgtgacatgc tgtttagcct gcccgagata aattctcatc
     4861 aaagtgccta ttgtccgcgc tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa
     4921 cgcgcctggc ggcaatggcc ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac
     4981 cgctgttgca tatctggctg ttaggcattc gtattgacgc caacgttatg caaggcatct
     5041 ggcaaatgac caaacagggc gatgcgataa cggggtcgat ggtctttttc tgcgttattg
     5101 gtgccccgct cattctggtg acctccatag cttatttatg gtttggtaac cgactgggaa
     5161 tgaatttacg tccggtactg ctgatgcttg agcgacttaa agagtgggta atgctcgata
     5221 tctacctggt cggcattggc gttgcttcta taaaggtaca ggattatgcc catatccagg
     5281 ctggtgtcgg cttgttctct tttgtggcgt tggtgatttt aacgacggtg acgttgtcac
     5341 atcttaatgt cgaagaactg tgggagcgat tttatccgca gcgccccgct acgcgtaggg
     5401 acgagaaact tcgtgtctgt cttgggtgcc attttaccgg ctatccagat cagcgtggtc
     5461 gctgcccgcg ttgccatatc ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct
     5521 gggcggcgct gttagcgtca atcgttttat tgttacctgc caacctgttg cctatttcta
     5581 tcatttatct gaacggagga cggcaggaag atacaattct ttccggaatt atgtcgctgg
     5641 caagtagcaa cattgcggtt gcaggaatcg tgtttatcgc cagtattctg gtaccgttta
     5701 ctaaagtgat cgtcatgttc actttactgt tgagcattca ttttaaatgc cagcaaggtt
     5761 tacgcacacg cattctgtta ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg
     5821 acctgtttgt catatcttta accatgtcgc tgattaatcg cgatcagatc ctcgctttta
     5881 ctatgggacc ggctgcgttt tatttcggcg cagcggtaat tttgactatt cttgctgtgg
     5941 aatggctgga cagccgctta ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg
     6001 actgaagcgc agattaaaaa taaacgccgt atctcacctt tctggctgct gccgttcatc
     6061 gcgctaatga ttgccagttg gctgatttgg gacagttatc aggaccgggg taataccgtc
     6121 accatcgact ttatgtcggc ggatggtatt gttccaggcc gtacgcctgt tcgttatcag
     6181 ggcgttgaag tcggaacagt gcaggatatc agcctcagcg acgatcttcg taagattgaa
     6241 gtcaaggtca gcatcaagtc cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg
     6301 ctggtgacgc caaaagcatc gttggcaggt gtctccgggc tggacgccct cgtcggtggg
     6361 aactatatcg gcatgatgcc gggtaaaggt aaagagcagg atcactttgt cgcactcgat
     6421 acccaaccga aatatcggct ggacaatggc gatctgatga tccacctgca agcccccgat
     6481 ctcggttcgc tgaacagcgg ttcattggtc tatttccgca agatcccggt gggaaaagtc
     6541 tacgactatg ccatcaatcc caacaagcaa ggcgtggtga ttgatgtcct gatcgagcgg
     6601 cgttttaccg atctggtgaa aaaaggtagc cgtttctgga acgtttccgg cgttgatgcc
     6661 aacgtcagta tcagtggcgc gaaggtgaaa ctggaaagcc tggcggcact ggttaacggt
     6721 gcgattgcct tcgattcacc agaagagtcg aaacctgccg aggcggaaga tacctttggt
     6781 ctgtatgaag atctggccca cagccagcgt ggcgtaataa taaaactgga actgccgagt
     6841 ggggccggat taaccgccga ctcgacgccg ttaatgtatc aggggctgga agtcggacag
     6901 ctgactaaac tggatttaaa tcctggtggt aaagtcacag gggaaatgac cgttgatccc
     6961 agcgtcgtta ccctgcttcg tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc
     7021 cttagcgatg ctaatctcag cgccctgctg accggcaaaa ccttcgagct ggtgcccggc
     7081 gatggcgagc cacgcaaaga gttcgttgtt gtgccaggcg aaaaagcact gctgcatgaa
     7141 cctgatgttc tgacgctgac cctgaccgcg ccggaaagtt acggtattga tgcgggtcag
     7201 ccgctcattc ttcacggcgt gcaggtaggc caggttatcg atcgtaaact caccagcaaa
     7261 ggcgtcacct ttaccgtcgc catcgagcct cagcatcgcg aactggtaaa aggcgatagc
     7321 aaatttgtcg tcaacagccg tgtcgatgtg aaggtggggc tggatggcgt tgagtttctc
     7381 ggtgccagcg cctcagaatg gatcaatggc gggatacgta ttctgccggg cgataaaggt
     7441 gagatgaaag ccagctatcc actgtatgcc aatttggaaa aagcgctgga gaacagcctt
     7501 agcgatttac ccaccacaac cgtgagtttg agtgcagaga cgctgccgga tgtgcaggca
     7561 ggatcggtag tgctgtaccg taaatttgaa gttggtgaag tgattaccgt gcgtccgcga
     7621 gctaacgcgt ttgatatcga tctgcatatt aagccggagt atcgcaacct tctgaccagc
     7681 aatagcgtgt tctgggcaga aggcggggcg aaagttcagc tgaatggtag tggtctgacc
     7741 gtacaggcat ccccgctctc cagagcatta aagggagcca ttagcttcga caacctcagc
     7801 ggtgccagcg ccagtcagcg taaaggcgat aagcgtattc tgtatgcttc cgaaacagcg
     7861 gcccgtgcgg ttggcgggca gattacgctt cacgctttcg atgccggaaa actggcggtc
     7921 gggatgccaa ttcgctatct cggtattgat atcgggcaaa tccagacgct ggatctgatt
     7981 accgcacgca atgaagtaca ggcaaaggcg gtactttacc cggaatatgt ccagaccttt
     8041 gctcgcggtg gtacgcgctt ctcagtggtc tccggcgttg atgccaacgt cagtatcagt
     8101 ggcgcgaagg tgaaactgga aagcctggcg gcactggtta acggttatat caacgtcgaa
     8161 ccaggccggg gcaatcctcg ccgcgacttt gaattacaag aggccaccat tactgattcg
     8221 cgttacctgg atggcttaag cattattgtt gaagcgccgg aagccggttc gttaggcatc
     8281 ggtacgcctg tgctgttccg tggtctggaa gtcggtacgg ttacaggaat gacgctgggg
     8341 acattgtcag atcgcgtgat gattgcgatg cgcatcagta aacgctatca acacctggtg
     8401 cgtaacaatt ccgtcttctg gttggcatcg ggttacagtc tggactttgg tctgacgggc
     8461 ggcgtagtga aaaccggcac ctttaaccag tttatccgtg gcggcatcgc cttcgccacg
     8521 cctccgggta cgccactggc accgaaagcc caggaaggca aacacttcct gttgcaggaa
     8581 agtgaaccga aagagtggcg tgaatgggga actgcgcttc ccaaataagt accaaattcc
     8641 agaaaagaga cgctgaaaag cgtctttttt cgttttggtc ct
//
