LOCUS       sequence-492986-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of of LetA with LetB(Y814B).
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  JOURNAL   Unpublished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            "Unpublished"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: pBEL3288
FEATURES             Location/Qualifiers
     source          1..8511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(262..1140)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1167..1451
                     /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     1374..1393
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1488..1505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1518..1538
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5536..5553)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5706..5792
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5884..5911
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5930..6021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6022..6090,6091..6882)
                     /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(6240..6259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6924..7379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7011..7030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7221..7242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      7490..8077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7978..7997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8231..8248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8414..8436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
       61 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
      121 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg
      181 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
      241 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
      301 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
      361 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
      421 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
      481 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
      541 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
      601 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
      661 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
      721 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
      781 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
      841 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
      901 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
      961 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     1021 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     1081 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     1141 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
     1201 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
     1261 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
     1321 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
     1381 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac
     1441 tgtttctcca tacccgtttt tttgggctag ccgaagggtg aattatgcat caccaccatc
     1501 accatcagca tcaacacgaa aatctgtatt ttcaggggat ggctcttaac acaccacaaa
     1561 ttacgccgac aaaaaagata acagtgaggg caatcggcga ggaactgccg cgtggtgatt
     1621 accaacgttg cccgcaatgt gacatgctgt ttagcctgcc cgagataaat tctcatcaaa
     1681 gtgcctattg tccgcgctgt caggcaaaaa ttcgtgacgg gcgcgactgg tcgctaacgc
     1741 gcctggcggc aatggccttc accatgctgt tgcttatgcc gtttgcctgg ggcgaaccgc
     1801 tgttgcatat ctggctgtta ggcattcgta ttgacgccaa cgttatgcaa ggcatctggc
     1861 aaatgaccaa acagggcgat gcgataacgg ggtcgatggt ctttttctgc gttattggtg
     1921 ccccgctcat tctggtgacc tccatagctt atttatggtt tggtaaccga ctgggaatga
     1981 atttacgtcc ggtactgctg atgcttgagc gacttaaaga gtgggtaatg ctcgatatct
     2041 acctggtcgg cattggcgtt gcttctataa aggtacagga ttatgcccat atccaggctg
     2101 gtgtcggctt gttctctttt gtggcgttgg tgattttaac gacggtgacg ttgtcacatc
     2161 ttaatgtcga agaactgtgg gagcgatttt atccgcagcg ccccgctacg cgtagggacg
     2221 agaaacttcg tgtctgtctt gggtgccatt ttaccggcta tccagatcag cgtggtcgct
     2281 gcccgcgttg ccatatcccg ctacgcctgc gtcgccgtca tagtctgcaa aaatgctggg
     2341 cggcgctgtt agcgtcaatc gttttattgt tacctgccaa cctgttgcct atttctatca
     2401 tttatctgaa cggaggacgg caggaagata caattctttc cggaattatg tcgctggcaa
     2461 gtagcaacat tgcggttgca ggaatcgtgt ttatcgccag tattctggta ccgtttacta
     2521 aagtgatcgt catgttcact ttactgttga gcattcattt taaatgccag caaggtttac
     2581 gcacacgcat tctgttactg cggatggtga cctggattgg tcgctggtcg atgctcgacc
     2641 tgtttgtcat atctttaacc atgtcgctga ttaatcgcga tcagatcctc gcttttacta
     2701 tgggaccggc tgcgttttat ttcggcgcag cggtaatttt gactattctt gctgtggaat
     2761 ggctggacag ccgcttactt tgggatgcac atgagtcagg aaacgcccgc ttcgacgact
     2821 gaagcgcaga ttaaaaataa acgccgtatc tcacctttct ggctgctgcc gttcatcgcg
     2881 ctaatgattg ccagttggct gatttgggac agttatcagg accggggtaa taccgtcacc
     2941 atcgacttta tgtcggcgga tggtattgtt ccaggccgta cgcctgttcg ttatcagggc
     3001 gttgaagtcg gaacagtgca ggatatcagc ctcagcgacg atcttcgtaa gattgaagtc
     3061 aaggtcagca tcaagtccga tatgaaagat gcgctgcgcg aagagactca gttctggctg
     3121 gtgacgccaa aagcatcgtt ggcaggtgtc tccgggctgg acgccctcgt cggtgggaac
     3181 tatatcggca tgatgccggg taaaggtaaa gagcaggatc actttgtcgc actcgatacc
     3241 caaccgaaat atcggctgga caatggcgat ctgatgatcc acctgcaagc ccccgatctc
     3301 ggttcgctga acagcggttc attggtctat ttccgcaaga tcccggtggg aaaagtctac
     3361 gactatgcca tcaatcccaa caagcaaggc gtggtgattg atgtcctgat cgagcggcgt
     3421 tttaccgatc tggtgaaaaa aggtagccgt ttctggaacg tttccggcgt tgatgccaac
     3481 gtcagtatca gtggcgcgaa ggtgaaactg gaaagcctgg cggcactggt taacggtgcg
     3541 attgccttcg attcaccaga agagtcgaaa cctgccgagg cggaagatac ctttggtctg
     3601 tatgaagatc tggcccacag ccagcgtggc gtaataataa aactggaact gccgagtggg
     3661 gccggattaa ccgccgactc gacgccgtta atgtatcagg ggctggaagt cggacagctg
     3721 actaaactgg atttaaatcc tggtggtaaa gtcacagggg aaatgaccgt tgatcccagc
     3781 gtcgttaccc tgcttcgtga aaatacccgc atcgaattac gcaacccgaa attatccctt
     3841 agcgatgcta atctcagcgc cctgctgacc ggcaaaacct tcgagctggt gcccggcgat
     3901 ggcgagccac gcaaagagtt cgttgttgtg ccaggcgaaa aagcactgct gcatgaacct
     3961 gatgttctga cgctgaccct gaccgcgccg gaaagttacg gtattgatgc gggtcagccg
     4021 ctcattcttc acggcgtgca ggtaggccag gttatcgatc gtaaactcac cagcaaaggc
     4081 gtcaccttta ccgtcgccat cgagcctcag catcgcgaac tggtaaaagg cgatagcaaa
     4141 tttgtcgtca acagccgtgt cgatgtgaag gtggggctgg atggcgttga gtttctcggt
     4201 gccagcgcct cagaatggat caatggcggg atacgtattc tgccgggcga taaaggtgag
     4261 atgaaagcca gctatccact gtatgccaat ttggaaaaag cgctggagaa cagccttagc
     4321 gatttaccca ccacaaccgt gagtttgagt gcagagacgc tgccggatgt gcaggcagga
     4381 tcggtagtgc tgtaccgtaa atttgaagtt ggtgaagtga ttaccgtgcg tccgcgagct
     4441 aacgcgtttg atatcgatct gcatattaag ccggagtatc gcaaccttct gaccagcaat
     4501 agcgtgttct gggcagaagg cggggcgaaa gttcagctga atggtagtgg tctgaccgta
     4561 caggcatccc cgctctccag agcattaaag ggagccatta gcttcgacaa cctcagcggt
     4621 gccagcgcca gtcagcgtaa aggcgataag cgtattctgt atgcttccga aacagcggcc
     4681 cgtgcggttg gcgggcagat tacgcttcac gctttcgatg ccggaaaact ggcggtcggg
     4741 atgccaattc gctatctcgg tattgatatc gggcaaatcc agacgctgga tctgattacc
     4801 gcacgcaatg aagtacaggc aaaggcggta ctttacccgg aatatgtcca gacctttgct
     4861 cgcggtggta cgcgcttctc agtggtcaca ccgcaaattt cggcagctgg cgttgagcat
     4921 cttgatacta tcctccagcc gtatatcaac gtcgaaccag gccggggcaa tcctcgccgc
     4981 gactttgaat tacaagaggc caccattact gattcgcgtt acctggatgg cttaagcatt
     5041 attgttgaag cgccggaagc cggttcgtta ggcatcggta cgcctgtgct gttccgtggt
     5101 ctggaagtcg gtacggttac aggaatgacg ctggggacat tgtcagatcg cgtgatgatt
     5161 gcgatgcgca tcagtaaacg ctatcaacac ctggtgcgta acaattccgt cttctggttg
     5221 gcatcgggtt agagtctgga ctttggtctg acgggcggcg tagtgaaaac cggcaccttt
     5281 aaccagttta tccgtggcgg catcgccttc gccacgcctc cgggtacgcc actggcaccg
     5341 aaagcccagg aaggcaaaca cttcctgttg caggaaagtg aaccgaaaga gtggcgtgaa
     5401 tggggaactg cgcttcccaa ataaggtacc aaattccaga aaagagacgc tgaaaagcgt
     5461 cttttttcgt tttggtcctg gtctgaggaa tggttgtaag cttggctgtt ttggcggatg
     5521 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     5581 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     5641 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
     5701 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     5761 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     5821 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     5881 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt
     5941 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6001 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6061 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6121 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6181 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6241 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6301 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6361 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6421 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6481 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6541 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     6601 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6661 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6721 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6781 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6841 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6901 tcatatatac tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7081 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     7141 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7201 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7261 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     7321 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta
     7381 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggctca gcagagcgca
     7621 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7921 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7981 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8041 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8101 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8161 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8221 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     8281 ccttacgcat ctgtgcggta tttcacaccg catagggtca tggctgcgcc ccgacacccg
     8341 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     8401 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     8461 gcgaggcaga aggagatggc gcccaacagt cccccggcca cggggcctgc c
//
