LOCUS       sequence-492970-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetB(R63K) with LetA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  TITLE     LetAB mutant plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7073)
  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            "Unpublished"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: pBEL3250
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             518..540
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(571..590)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(572..590)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1099..1290
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1284..1306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1392..1531
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1447..1466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1546..1563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1717..2305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1797..1816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2476..3267,3268..3336))
                     /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     3099..3118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3337..3441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3509..3527
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3572..3619)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3615..3633
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gaccaggtag atatcgagca ttacccactc tttaagtcgc tcaagcatca gcagtaccgg
       61 acgtaaattc attcccagtc ggttaccaaa ccataaataa gctatggagg tcaccagaat
      121 gagcggggca ccaataacgc agaaaaagac catcgacccc gttatcgcat cgccctgttt
      181 ggtcatttgc cagatgcctt gcataacgtt ggcgtcaata cgaatgccta acagccagat
      241 atgcaacagc ggttcgcccc aggcaaacgg cataagcaac agcatggtga aggccattgc
      301 cgccaggcgc gttagcgacc agtcgcgccc gtcacgaatt tttgcctgac agcgcggaca
      361 ataggcactt tgatgagaat ttatctcggg caggctaaac agcatgtcac attgcgggca
      421 acgttggtaa tcaccacgcg gcagttcctc gccgattgcc ctcactgtta tcttttttgt
      481 cggcgtaatt tgtggtgtgt taagagccat atgtatatct ccttcttaaa gttaaacaaa
      541 attatttcta gagggaaacc gttgtggtct ccctatagtg agtcgtatta atttcgcggg
      601 atcgagatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
      661 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
      721 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
      781 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
      841 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
      901 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
      961 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     1021 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     1081 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     1141 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     1201 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     1261 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1321 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1381 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     1441 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
     1501 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     1561 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1621 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1681 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1741 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1801 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1861 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1921 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1981 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2041 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2101 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2161 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2221 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2281 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2341 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2401 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2461 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2521 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2581 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2641 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2701 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2761 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     2821 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2881 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2941 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3001 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3061 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     3121 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3181 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3241 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3301 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3361 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3421 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3481 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     3541 agaattatcc ggatatagtt cctcctttca gcaaaaaacc cctcaagacc cgtttagagg
     3601 ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc
     3661 gggctttgtt agcagccgga tctgctcgag gtgggcatta tttgggaagc gcagttcccc
     3721 attcacgcca ctctttcggt tcactttcct gcaacaggaa gtgtttgcct tcctgggctt
     3781 tcggtgccag tggcgtaccc ggaggcgtgg cgaaggcgat gccgccacgg ataaactggt
     3841 taaaggtgcc ggttttcact acgccgcccg tcagaccaaa gtccagactg taacccgatg
     3901 ccaaccagaa gacggaattg ttacgcacca ggtgttgata gcgtttactg atgcgcatcg
     3961 caatcatcac gcgatctgac aatgtcccca gcgtcattcc tgtaaccgta ccgacttcca
     4021 gaccacggaa cagcacaggc gtaccgatgc ctaacgaacc ggcttccggc gcttcaacaa
     4081 taatgcttaa gccatccagg taccgcgaat cagtaatggt ggcctcttgt aattcaaagt
     4141 cgcggcgagg attgccccgg cctggttcga cgttgatata cggctggagg atagtatcaa
     4201 gatgctcaac gccagctgcc gaaatttgcg gtgtgaccac tgagaagcgc gtaccaccgc
     4261 gagcaaaggt ctggacatat tccgggtaaa gtaccgcctt tgcctgtact tcattgcgtg
     4321 cggtaatcag atccagcgtc tggatttgcc cgatgtcaat accgagatag cgaattggca
     4381 tcccgaccgc cagttttccg gcatcgaaag cgtgaagcgt aatctgcccg ccaaccgcac
     4441 gggccgctgt ttcggaagca tacagaatac gcttatcgcc tttacgctga ctggcgctgg
     4501 caccgctgag gttgtcgaag ctaatggctc cctttagagc tctggagagc ggggatgcct
     4561 gtacggtcag accactacca ttcagctgaa ctttcgcccc gccttctgcc cagaacacgc
     4621 tattgctggt cagaaggttg cgatactccg gcttaatatg cagatcgatg tcaaacgcgt
     4681 tagctcgcgg acgcacggta atcacttcac caacttcaaa tttacggtac agcactaccg
     4741 atcctgcctg cacatccggc agcgtctctg cactcaaact cacggttgtg gtgggtaaat
     4801 cgctaaggct gttctccagc gctttttcca aattggcata cagtggatag ctggctttca
     4861 tctcaccttt atcgcccggc agaatacgga tcccgccatt gatccattct gaggcgctgg
     4921 caccgagaaa ctcaacgcca tccagcccca ccttcacatc gacacggctg ttgacgacaa
     4981 atttgctatc gccttttacc agttcgcgat gctgaggctc gatggcgacg gtaaaggtga
     5041 cgcctttgct ggtgagttta cgatcgataa cctggcctac ctgcacgccg tgaagaatga
     5101 gcggctgacc cgcatcaata ccgtaacttt ccggcgcggt cagggtcagc gtcagaacat
     5161 caggttcatg cagcagtgct ttttcgcctg gcacaacaac gaattctttg cgtggctcgc
     5221 catcgccggg caccagctcg aaggttttgc cggtcagcag ggcgctgaga ttagcatcgc
     5281 taagggataa tttcgggttg cgtaattcga tgcgggtatt ttcacgaagc agggtaacga
     5341 cgctgggatc aacggtcatt tcccctgtga ctttaccacc aggatttaaa tccagtttag
     5401 tcagctgtcc gacttccagc ccctgataca ttaacggcgt cgagtcggcg gttaatccgg
     5461 ccccactcgg cagttccagt tttattatta cgccacgctg gctgtgggcc agatcttcat
     5521 acagaccaaa ggtatcttcc gcctcggcag gtttcgactc ttctggtgaa tcgaaggcaa
     5581 tcgcaccgtt aaccagtgcc gccaagcttt ccagtttcac cttcgcgcca ctgatactga
     5641 cgttggcatc aacgccggaa acgttccaga aacggctacc ttttttcacc agatcggtaa
     5701 aacgccgctc gatcaggaca tcaatcacca cgccttgctt gttgggattg atggcatagt
     5761 cgtagacttt tcccaccggg atcttgcgga aatagaccaa tgaaccgctg ttcagcgaac
     5821 cgagatcggg ggcttgcagg tggatcatca gatcgccatt gtccagccga tatttcggtt
     5881 gggtatcgag tgcgacaaag tgatcctgct ctttaccttt cccgggcatc atgccgatat
     5941 agttcccacc gacgagggcg tccagcccgg agacacctgc caacgatgct tttggcgtca
     6001 ccagccagaa ctgagtctct tcgcgcagcg catctttcat atcggacttg atgctgacct
     6061 tgacttcaat cttacgaaga tcgtcgctga ggctgatgtc ctgcactgtt ccgacttcaa
     6121 cgccctgata acgaacaggc gtcttgcctg gaacaatacc atccgccgac ataaagtcga
     6181 tggtgacggt attaccccgg tcctgataac tgtcccaaat cagccaactg gcaatcatta
     6241 gcgcgatgaa cggcagcagc cagaaaggtg agatacggcg tttattttta atctgcgctt
     6301 cagtcgtcga agcgggcgtt tcctgactca tgtgcatccc aaagtaagcg gctgtccagc
     6361 cattccacag caagaatagt caaaattacc gctgcgccga aataaaacgc agccggtccc
     6421 atagtaaaag cgaggatctg atcgcgatta atcagcgaca tggttaaaga tatgacaaac
     6481 aggtcgagca tcgaccagcg accaatccag gtcaccatcc gcagtaacag aatgcgtgtg
     6541 cgtaaacctt gctggcattt aaaatgaatg ctcaacagta aagtgaacat gacgatcact
     6601 ttagtaaacg ggaccagaat actggcgata aacacgattc ctgcaaccgc aatgttgcta
     6661 cttgccagcg acataattcc ggaaagaatt gtatcttcct gccgtcctcc gttcagataa
     6721 atgatagaaa taggcaacag gttggcaggt aacaataaaa cgattgacgc taacagcgcc
     6781 gcccagcatt tttgcagact atgacggcga cgcaggcgta gcgggatatg gcaacgcggg
     6841 cagcgaccac gctgatctgg atagccggta aaatggcacc ctaggcagac acgaagtttc
     6901 tcgtccctac gcgtagcggg gcgctgcgga taaaatcgct cccacagttc ttcgacatta
     6961 agatgtgaca acgtcaccgt cgttaaaatc accaacgcca caaaagagaa caagccgaca
     7021 ccagcctgga tatgggcata atcctgtacc tttatagaag caacgccaat gcc
//
