LOCUS       sequence-491674-        8857 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Expresses the fusion of TadDE (dual deaminase) and T7RNAP for 
            gene-specific mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8857)
  TITLE     TadDE-T7RNAP expression plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBH001 (TadDE-T7RNAP)
FEATURES             Location/Qualifiers
     source          1..8857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4598..4852
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     primer_bind     complement(4891..4908)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5061..5147
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5239..5266
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5285..5376
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5834..6289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5921..5940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6131..6152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6845..7504)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7419..7438
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7505..7607)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7999..8016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8133..8678)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(8213..8232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8754..8773
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcgaatt cgagctcagg
     1321 aggaattcac catgagtgaa gttgaattca gccatgaata ttggatgcgt catgccctga
     1381 ccctggcaaa acgtgcccgc gatgaaggtg aagctccggt tggtgcggtg ctggttttaa
     1441 ataatcgtgt gattggcgaa ggctggaatc gtcgtattgg tctgcatgat cctaccgcac
     1501 atgcagaaat catggccctg cgtcagggtg gtctggttat gcaaaattct cgcctgattg
     1561 atgccaccct gtatgtgacc tttgaacctt gtgtgatgtg tgcgggtgca atgattaata
     1621 gtcgtattgg ccgtgttgtg tttggcgttc gtaatagcaa gcgcggtgcc gccggcagcc
     1681 tgatgaatgt tctgaattat cctggcatga atcatcgcgt ggaaattacc gagggtattc
     1741 tggccgatga atgtgccgca ctgctgtgcg atttttatcg catgccgcgt caagtgttta
     1801 atgcacaaaa gaaagcacag agcagcatta atagcggcag cgagactccc gggacctcag
     1861 agtccgccac acccgaaagt aacacgatta acatcgctaa gaacgacttc tctgacatcg
     1921 aactggctgc tatcccgttc aacactctgg ctgaccatta cggtgagcgt ttagctcgcg
     1981 aacagttggc ccttgagcat gagtcttacg agatgggtga agcacgcttc cgcaagatgt
     2041 ttgagcgtca acttaaagct ggtgaggttg cggataacgc tgccgccaag cctctcatca
     2101 ctaccctact ccctaagatg attgcacgca tcaacgactg gtttgaggaa gtgaaagcta
     2161 agcgcggcaa gcgcccgaca gccttccagt tcctgcaaga aatcaagccg gaagccgtag
     2221 cgtacatcac cattaagacc actctggctt gcctaaccag tgctgacaat acaaccgttc
     2281 aggctgtagc aagcgcaatc ggtcgggcca ttgaggacga ggctcgcttc ggtcgtatcc
     2341 gtgaccttga agctaagcac ttcaagaaaa acgttgagga acaactcaac aagcgcgtag
     2401 ggcacgtcta caagaaagca tttatgcaag ttgtcgaggc tgacatgctc tctaagggtc
     2461 tactcggtgg cgaggcgtgg tcttcgtggc ataaggaaga ctctattcat gtaggagtac
     2521 gctgcatcga gatgctcatt gagtcaaccg gaatggttag cttacaccgc caaaatgctg
     2581 gcgtagtagg tcaagactct gagactatcg aactcgcacc tgaatacgct gaggctatcg
     2641 caacccgtgc aggtgcgctg gctggcatct ctccgatgtt ccaaccttgc gtagttcctc
     2701 ctaagccgtg gactggcatt actggtggtg gctattgggc taacggtcgt cgtcctctgg
     2761 cgctggtgcg tactcacagt aagaaagcac tgatgcgcta cgaagacgtt tacatgcctg
     2821 aggtgtacaa agcgattaac attgcgcaaa acaccgcatg gaaaatcaac aagaaagtcc
     2881 tagcggtcgc caacgtaatc accaagtgga agcattgtcc ggtcgaggac atccctgcga
     2941 ttgagcgtga agaactcccg atgaaaccgg aagacatcga catgaatcct gaggctctca
     3001 ccgcgtggaa acgtgctgcc gctgctgtgt accgcaagga caaggctcgc aagtctcgcc
     3061 gtatcagcct tgagttcatg cttgagcaag ccaataagtt tgctaaccat aaggccatct
     3121 ggttccctta caacatggac tggcgcggtc gtgtttacgc tgtgtcaatg ttcaacccgc
     3181 aaggtaacga tatgaccaaa ggactgctta cgctggcgaa aggtaaacca atcggtaagg
     3241 aaggttacta ctggctgaaa atccacggtg caaactgtgc gggtgtcgat aaggttccgt
     3301 tccctgagcg catcaagttc attgaggaaa accacgagaa catcatggct tgcgctaagt
     3361 ctccactgga gaacacttgg tgggctgagc aagattctcc gttctgcttc cttgcgttct
     3421 gctttgagta cgctggggta cagcaccacg gcctgagcta taactgctcc cttccgctgg
     3481 cgtttgacgg gtcttgctct ggcatccagc acttctccgc gatgctccga gatgaggtag
     3541 gtggtcgcgc ggttaacttg cttcctagtg aaaccgttca ggacatctac gggattgttg
     3601 ctaagaaagt caacgagatt ctacaagcag acgcaatcaa tgggaccgat aacgaagtag
     3661 ttaccgtgac cgatgagaac actggtgaaa tctctgagaa agtcaagctg ggcactaagg
     3721 cactggctgg tcaatggctg gcttacggtg ttactcgcag tgtgactaag cgttcagtca
     3781 tgacgctggc ttacgggtcc aaagagttcg gcttccgtca acaagtgctg gaagatacca
     3841 ttcagccagc tattgattcc ggcaagggtc tgatgttcac tcagccgaat caggctgctg
     3901 gatacatggc taagctgatt tgggaatctg tgagcgtgac ggtggtagct gcggttgaag
     3961 caatgaactg gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa gataagaaga
     4021 ctggagagat tcttcgcaag cgttgcgctg tgcattgggt aactcctgat ggtttccctg
     4081 tgtggcagga atacaagaag cctattcaga cgcgcttgaa cctgatgttc ctcggtcagt
     4141 tccgcttaca gcctaccatt aacaccaaca aagatagcga gattgatgca cacaaacagg
     4201 agtctggtat cgctcctaac tttgtacaca gccaagacgg tagccacctt cgtaagactg
     4261 tagtgtgggc acacgagaag tacggaatcg aatcttttgc actgattcac gactccttcg
     4321 gtaccattcc ggctgacgct gcgaacctgt tcaaagcagt gcgcgaaact atggttgaca
     4381 cctatgagtc ttgtgatgta ctggctgatt tctacgacca gttcgctgac cagttgcacg
     4441 agtctcaatt ggacaaaatg ccagcacttc cggctaaagg taacttgaac ctccgtgaca
     4501 tcttagagtc ggacttcgcg ttcgcgtaac tgcagcccgg ggatcctaac agagcgcgct
     4561 ctgtttgagt actagcaata aataaggagg attttttatg accaaccttt ccgacatcat
     4621 agagaaggaa acaggcaaac agttggtcat ccaagagtcg atactcatgc ttcctgaaga
     4681 agttgaggag gtcattggga ataagccgga aagtgacatt ctcgtacaca ctgcgtatga
     4741 tgagagcacc gatgagaacg tgatgctgct cacgtcagat gccccagagt acaaaccctg
     4801 ggctctggtg attcaggact ctaatggaga gaacaagatc aagatgctgt aaaagcttgg
     4861 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     4921 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     4981 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     5041 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     5101 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     5161 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     5221 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     5281 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5341 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5461 gtgaaagtaa aagatgctga agatcagttg ggtgcagcaa actattaact ggcgaactac
     5521 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5581 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5641 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5701 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5761 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5821 tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5881 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     6061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     6181 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6241 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta
     6301 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6541 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6601 tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca cacggtcaca
     6661 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     6721 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     6781 tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa
     6841 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     6901 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     6961 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     7021 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     7081 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     7141 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     7201 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     7261 gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa
     7321 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     7381 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     7441 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     7501 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     7561 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     7621 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     7681 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
     7741 acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc
     7801 agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga
     7861 catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg
     7921 aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata
     7981 caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     8041 agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa
     8101 cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag
     8161 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     8221 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     8281 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     8341 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     8401 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     8461 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     8521 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     8581 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     8641 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga
     8701 taaaatattt ctaggctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     8761 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     8821 ccggcgtaga ggatctgctc atgtttgaca gcttatc
//
