LOCUS       sequence-492987-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of of LetA with LetB(F833B).
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  JOURNAL   Unpublished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            "Unpublished"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: pBEL2637
FEATURES             Location/Qualifiers
     source          1..8511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..100)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(266..283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(437..1024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(517..536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(1135..1590)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1272..1293)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1484..1503
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(1632..2423,2424..2492))
                     /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     2255..2274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2493..2584)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2603..2630)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2722..2808)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2961..2978
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(6976..6996)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(7009..7026)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(7063..7347)
                     /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     complement(7121..7140)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             7374..8252
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(8432..8451)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttctgcct cgcgcgtttc
       61 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
      121 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
      181 cggggcgcag ccatgaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      241 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      301 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      361 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      421 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      841 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
      901 tgctgagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaattg
     1141 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1201 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt
     1261 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1321 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     1381 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1441 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1501 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     1561 ccgcgcttaa tgcgccgcta cagggcgcgt aaatcaatct aaagtatata tgagtaaact
     1621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1921 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     1981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2221 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     2281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2581 caaaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttaatttg
     2641 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcgcaac
     2701 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     2761 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggcagt
     2821 tccctactct cgcatgggga gaccccacac taccatcggc gctacggcgt ttcacttctg
     2881 agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa ttctgtttta
     2941 tcagaccgct tctgcgttct gatttaatct gtatcaggct gaaaatcttc tctcatccgc
     3001 caaaacagcc aagcttacaa ccattcctca gaccaggacc aaaacgaaaa aagacgcttt
     3061 tcagcgtctc ttttctggaa tttggtacct tatttgggaa gcgcagttcc ccattcacgc
     3121 cactctttcg gttcactttc ctgcaacagg aagtgtttgc cttcctgggc tttcggtgcc
     3181 agtggcgtac ccggaggcgt ggcgaaggcg atgccgccac ggatctactg gttaaaggtg
     3241 ccggttttca ctacgccgcc cgtcagacca aagtccagac tgtaacccga tgccaaccag
     3301 aagacggaat tgttacgcac caggtgttga tagcgtttac tgatgcgcat cgcaatcatc
     3361 acgcgatctg acaatgtccc cagcgtcatt cctgtaaccg taccgacttc cagaccacgg
     3421 aacagcacag gcgtaccgat gcctaacgaa ccggcttccg gcgcttcaac aataatgctt
     3481 aagccatcca ggtaacgcga atcagtaatg gtggcctctt gtaattcaaa gtcgcggcga
     3541 ggattgcccc ggcctggttc gacgttgata tacggctgga ggatagtatc aagatgctca
     3601 acgccagctg ccgaaatttg cggtgtgacc actgagaagc gcgtaccacc gcgagcaaag
     3661 gtctggacat attccgggta aagtaccgcc tttgcctgta cttcattgcg tgcggtaatc
     3721 agatccagcg tctggatttg cccgatatca ataccgagat agcgaattgg catcccgacc
     3781 gccagttttc cggcatcgaa agcgtgaagc gtaatctgcc cgccaaccgc acgggccgct
     3841 gtttcggaag catacagaat acgcttatcg cctttacgct gactggcgct ggcaccgctg
     3901 aggttgtcga agctaatggc tccctttaat gctctggaga gcggggatgc ctgtacggtc
     3961 agaccactac cattcagctg aactttcgcc ccgccttctg cccagaacac gctattgctg
     4021 gtcagaaggt tgcgatactc cggcttaata tgcagatcga tatcaaacgc gttagctcgc
     4081 ggacgcacgg taatcacttc accaacttca aatttacggt acagcactac cgatcctgcc
     4141 tgcacatccg gcagcgtctc tgcactcaaa ctcacggttg tggtgggtaa atcgctaagg
     4201 ctgttctcca gcgctttttc caaattggca tacagtggat agctggcttt catctcacct
     4261 ttatcgcccg gcagaatacg tatcccgcca ttgatccatt ctgaggcgct ggcaccgaga
     4321 aactcaacgc catccagccc caccttcaca tcgacacggc tgttgacgac aaatttgcta
     4381 tcgcctttta ccagttcgcg atgctgaggc tcgatggcga cggtaaaggt gacgcctttg
     4441 ctggtgagtt tacgatcgat aacctggcct acctgcacgc cgtgaagaat gagcggctga
     4501 cccgcatcaa taccgtaact ttccggcgcg gtcagggtca gcgtcagaac atcaggttca
     4561 tgcagcagtg ctttttcgcc tggcacaaca acgaactctt tgcgtggctc gccatcgccg
     4621 ggcaccagct cgaaggtttt gccggtcagc agggcgctga gattagcatc gctaagggat
     4681 aatttcgggt tgcgtaattc gatgcgggta ttttcacgaa gcagggtaac gacgctggga
     4741 tcaacggtca tttcccctgt gactttacca ccaggattta aatccagttt agtcagctgt
     4801 ccgacttcca gcccctgata cattaacggc gtcgagtcgg cggttaatcc ggccccactc
     4861 ggcagttcca gttttattat tacgccacgc tggctgtggg ccagatcttc atacagacca
     4921 aaggtatctt ccgcctcggc aggtttcgac tcttctggtg aatcgaaggc aatcgcaccg
     4981 ttaaccagtg ccgccaggct ttccagtttc accttcgcgc cactgatact gacgttggca
     5041 tcaacgccgg aaacgttcca gaaacggcta ccttttttca ccagatcggt aaaacgccgc
     5101 tcgatcagga catcaatcac cacgccttgc ttgttgggat tgatggcata gtcgtagact
     5161 tttcccaccg ggatcttgcg gaaatagacc aatgaaccgc tgttcagcga accgagatcg
     5221 ggggcttgca ggtggatcat cagatcgcca ttgtccagcc gatatttcgg ttgggtatcg
     5281 agtgcgacaa agtgatcctg ctctttacct ttacccggca tcatgccgat atagttccca
     5341 ccgacgaggg cgtccagccc ggagacacct gccaacgatg cttttggcgt caccagccag
     5401 aactgagtct cttcgcgcag cgcatctttc atatcggact tgatgctgac cttgacttca
     5461 atcttacgaa gatcgtcgct gaggctgata tcctgcactg ttccgacttc aacgccctga
     5521 taacgaacag gcgtacggcc tggaacaata ccatccgccg acataaagtc gatggtgacg
     5581 gtattacccc ggtcctgata actgtcccaa atcagccaac tggcaatcat tagcgcgatg
     5641 aacggcagca gccagaaagg tgagatacgg cgtttatttt taatctgcgc ttcagtcgtc
     5701 gaagcgggcg tttcctgact catgtgcatc ccaaagtaag cggctgtcca gccattccac
     5761 agcaagaata gtcaaaatta ccgctgcgcc gaaataaaac gcagccggtc ccatagtaaa
     5821 agcgaggatc tgatcgcgat taatcagcga catggttaaa gatatgacaa acaggtcgag
     5881 catcgaccag cgaccaatcc aggtcaccat ccgcagtaac agaatgcgtg tgcgtaaacc
     5941 ttgctggcat ttaaaatgaa tgctcaacag taaagtgaac atgacgatca ctttagtaaa
     6001 cggtaccaga atactggcga taaacacgat tcctgcaacc gcaatgttgc tacttgccag
     6061 cgacataatt ccggaaagaa ttgtatcttc ctgccgtcct ccgttcagat aaatgataga
     6121 aataggcaac aggttggcag gtaacaataa aacgattgac gctaacagcg ccgcccagca
     6181 tttttgcaga ctatgacggc gacgcaggcg tagcgggata tggcaacgcg ggcagcgacc
     6241 acgctgatct ggatagccgg taaaatggca cccaagacag acacgaagtt tctcgtccct
     6301 acgcgtagcg gggcgctgcg gataaaatcg ctcccacagt tcttcgacat taagatgtga
     6361 caacgtcacc gtcgttaaaa tcaccaacgc cacaaaagag aacaagccga caccagcctg
     6421 gatatgggca taatcctgta cctttataga agcaacgcca atgccgacca ggtagatatc
     6481 gagcattacc cactctttaa gtcgctcaag catcagcagt accggacgta aattcattcc
     6541 cagtcggtta ccaaaccata aataagctat ggaggtcacc agaatgagcg gggcaccaat
     6601 aacgcagaaa aagaccatcg accccgttat cgcatcgccc tgtttggtca tttgccagat
     6661 gccttgcata acgttggcgt caatacgaat gcctaacagc cagatatgca acagcggttc
     6721 gccccaggca aacggcataa gcaacagcat ggtgaaggcc attgccgcca ggcgcgttag
     6781 cgaccagtcg cgcccgtcac gaatttttgc ctgacagcgc ggacaatagg cactttgatg
     6841 agaatttatc tcgggcaggc taaacagcat gtcacattgc gggcaacgtt ggtaatcacc
     6901 acgcggcagt tcctcgccga ttgccctcac tgttatcttt tttgtcggcg taatttgtgg
     6961 tgtgttaaga gccatcccct gaaaatacag attttcgtgt tgatgctgat ggtgatggtg
     7021 gtgatgcata attcaccctt cggctagccc aaaaaaacgg gtatggagaa acagtagaga
     7081 gttgcgataa aaagcgtcag gtaggatccg ctaatcttat ggataaaaat gctatggcat
     7141 agcaaagtgt gacgccgtgc aaataatcaa tgtggacttt tctgccgtga ttatagacac
     7201 ttttgttacg cgtttttgtc atggctttgg tcccgctttg ttacagaatg cttttaataa
     7261 gcggggttac cggtttggtt agcgagaaga gccagtaaaa gacgcagtga cggcaatgtc
     7321 tgatgcaata tggacaattg gtttcttctc tgaatggcgg gagtatgaaa agtatggctg
     7381 aagcgcaaaa tgatcccctg ctgccgggat actcgtttaa tgcccatctg gtggcgggtt
     7441 taacgccgat tgaggccaac ggttatctcg atttttttat cgaccgaccg ctgggaatga
     7501 aaggttatat tctcaatctc accattcgcg gtcagggggt ggtgaaaaat cagggacgag
     7561 aatttgtttg ccgaccgggt gatattttgc tgttcccgcc aggagagatt catcactacg
     7621 gtcgtcatcc ggaggctcgc gaatggtatc accagtgggt ttactttcgt ccgcgcgcct
     7681 actggcatga atggcttaac tggccgtcaa tatttgccaa tacggggttc tttcgcccgg
     7741 atgaagcgca ccagccgcat ttcagcgacc tgtttgggca aatcattaac gccgggcaag
     7801 gggaagggcg ctattcggag ctgctggcga taaatctgct tgagcaattg ttactgcggc
     7861 gcatggaagc gattaacgag tcgctccatc caccgatgga taatcgggta cgcgaggctt
     7921 gtcagtacat cagcgatcac ctggcagaca gcaattttga tatcgccagc gtcgcacagc
     7981 atgtttgctt gtcgccgtcg cgtctgtcac atcttttccg ccagcagtta gggattagcg
     8041 tcttaagctg gcgcgaggac caacgtatca gccaggcgaa gctgcttttg agcaccaccc
     8101 ggatgcctat cgccaccgtc ggtcgcaatg ttggttttga cgatcaactc tatttctcgc
     8161 gggtatttaa aaaatgcacc ggggccagcc cgagcgagtt ccgtgccggt tgtgaagaaa
     8221 aagtgaatga tgtagccgtc aagttgtcat aattggtaac gaatcagaca attgacggct
     8281 tgacggagta gcatagggtt tgcagaatcc ctgcttcgtc catttgacag gcacattatg
     8341 catcgatgat aagctgtcaa acatgagcag atcctctacg ccggacgcat cgtggccggc
     8401 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     8461 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat g
//
