LOCUS       sequence-492988-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of of LetA with LetB(W476B).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8511)
  TITLE     LetAB mutant plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8511)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBEL3287
FEATURES             Location/Qualifiers
     source          1..8511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        683..967
                     /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     890..909
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1004..1021
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1034..1054
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5052..5069)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5222..5308
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5400..5427
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5446..5537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5538..5606,5607..6398)
                     /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(5756..5775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6440..6895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6527..6546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6737..6758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      7006..7593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7494..7513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7747..7764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7930..7952
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8090..8109
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(8289..8511,1..656))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
       61 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
      121 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac
      181 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
      241 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
      301 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
      361 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
      421 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
      481 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
      541 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
      601 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
      661 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
      721 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
      781 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat
      841 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca
      901 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt
      961 tctccatacc cgtttttttg ggctagccga agggtgaatt atgcatcacc accatcacca
     1021 tcagcatcaa cacgaaaatc tgtattttca ggggatggct cttaacacac cacaaattac
     1081 gccgacaaaa aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca
     1141 acgttgcccg caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc
     1201 ctattgtccg cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct
     1261 ggcggcaatg gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt
     1321 gcatatctgg ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat
     1381 gaccaaacag ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc
     1441 gctcattctg gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt
     1501 acgtccggta ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct
     1561 ggtcggcatt ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt
     1621 cggcttgttc tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa
     1681 tgtcgaagaa ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa
     1741 acttcgtgtc tgtcttgggt gccattttac cggctatcca gatcagcgtg gtcgctgccc
     1801 gcgttgccat atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc
     1861 gctgttagcg tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta
     1921 tctgaacgga ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag
     1981 caacattgcg gttgcaggaa tcgtgtttat cgccagtatt ctggtaccgt ttactaaagt
     2041 gatcgtcatg ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac
     2101 acgcattctg ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt
     2161 tgtcatatct ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg
     2221 accggctgcg ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct
     2281 ggacagccgc ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag
     2341 cgcagattaa aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa
     2401 tgattgccag ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg
     2461 actttatgtc ggcggatggt attgttccag gccgtacgcc tgttcgttat cagggcgttg
     2521 aagtcggaac agtgcaggat atcagcctca gcgacgatct tcgtaagatt gaagtcaagg
     2581 tcagcatcaa gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga
     2641 cgccaaaagc atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata
     2701 tcggcatgat gccgggtaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac
     2761 cgaaatatcg gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt
     2821 cgctgaacag cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact
     2881 atgccatcaa tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta
     2941 ccgatctggt gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca
     3001 gtatcagtgg cgcgaaggtg aaactggaaa gcctggcggc actggttaac ggtgcgattg
     3061 ccttcgattc accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg
     3121 aagatctggc ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg
     3181 gattaaccgc cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta
     3241 aactggattt aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg
     3301 ttaccctgct tcgtgaaaat acccgcatcg aattacgcaa cccgaaatta tcccttagcg
     3361 atgctaatct cagcgccctg ctgaccggca aaaccttcga gctggtgccc ggcgatggcg
     3421 agccacgcaa agagttcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg
     3481 ttctgacgct gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca
     3541 ttcttcacgg cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca
     3601 cctttaccgt cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg
     3661 tcgtcaacag ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca
     3721 gcgcctcaga atagatcaat ggcgggatac gtattctgcc gggcgataaa ggtgagatga
     3781 aagccagcta tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt
     3841 tacccaccac aaccgtgagt ttgagtgcag agacgctgcc ggatgtgcag gcaggatcgg
     3901 tagtgctgta ccgtaaattt gaagttggtg aagtgattac cgtgcgtccg cgagctaacg
     3961 cgtttgatat cgatctgcat attaagccgg agtatcgcaa ccttctgacc agcaatagcg
     4021 tgttctgggc agaaggcggg gcgaaagttc agctgaatgg tagtggtctg accgtacagg
     4081 catccccgct ctccagagca ttaaagggag ccattagctt cgacaacctc agcggtgcca
     4141 gcgccagtca gcgtaaaggc gataagcgta ttctgtatgc ttccgaaaca gcggcccgtg
     4201 cggttggcgg gcagattacg cttcacgctt tcgatgccgg aaaactggcg gtcgggatgc
     4261 caattcgcta tctcggtatt gatatcgggc aaatccagac gctggatctg attaccgcac
     4321 gcaatgaagt acaggcaaag gcggtacttt acccggaata tgtccagacc tttgctcgcg
     4381 gtggtacgcg cttctcagtg gtcacaccgc aaatttcggc agctggcgtt gagcatcttg
     4441 atactatcct ccagccgtat atcaacgtcg aaccaggccg gggcaatcct cgccgcgact
     4501 ttgaattaca agaggccacc attactgatt cgcgttacct ggatggctta agcattattg
     4561 ttgaagcgcc ggaagccggt tcgttaggca tcggtacgcc tgtgctgttc cgtggtctgg
     4621 aagtcggtac ggttacagga atgacgctgg ggacattgtc agatcgcgtg atgattgcga
     4681 tgcgcatcag taaacgctat caacacctgg tgcgtaacaa ttccgtcttc tggttggcat
     4741 cgggttacag tctggacttt ggtctgacgg gcggcgtagt gaaaaccggc acctttaacc
     4801 agtttatccg tggcggcatc gccttcgcca cgcctccggg tacgccactg gcaccgaaag
     4861 cccaggaagg caaacacttc ctgttgcagg aaagtgaacc gaaagagtgg cgtgaatggg
     4921 gaactgcgct tcccaaataa ggtaccaaat tccagaaaag agacgctgaa aagcgtcttt
     4981 tttcgttttg gtcctggtct gaggaatggt tgtaagcttg gctgttttgg cggatgagag
     5041 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     5101 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     5161 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     5221 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     5281 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     5341 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     5401 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     5461 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5521 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5581 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5641 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5701 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5761 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5821 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5881 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5941 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6001 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6061 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6121 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6181 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6241 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6301 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6361 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6421 atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6481 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6541 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6601 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     6661 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6721 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6781 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6841 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag
     6901 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6961 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7021 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7081 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggctcagcag agcgcagata
     7141 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7681 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7741 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     7801 acgcatctgt gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa
     7861 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     7921 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     7981 ggcagaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     8041 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     8101 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     8161 cgtagaggat ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg
     8221 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
     8281 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     8341 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     8401 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     8461 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc t
//
