LOCUS       sequence-97715-e        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses both fragments of T7 RNAP split at position 179. Both 
            fragments are driven by PBAD. The C-terminal fragment has the point 
            mutation R756K. Contains a constitutive araC ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  AUTHORS   Shis DL, Bennett MR
  TITLE     Library of synthetic transcriptional AND gates built with split T7 
            RNA polymerase mutants.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5028-33. doi: 
            10.1073/pnas.1220157110. Epub 2013 Mar 11.
  PUBMED    23479654
REFERENCE   2  (bases 1 to 7682)
  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"; doi: 
            "10.1073/pnas.1220157110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-03-26- 26"; volume: "110"; issue: "13"; pages: "5028-33"
COMMENT     Sequence Label: pTSara-R756K
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(288..315)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3725..3742)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3895..3981
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4073..4100
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4119..4210
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4664..5119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4751..4770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(5674..6333)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6248..6267
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6334..6436)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6828..6845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6962..7507)
                     /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(7042..7061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7579..7598
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaacagac attgccgtca ctgcgtcttt
      121 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
      181 agcgggacca aagccatgac aaaaacgcgt aatttcaaca cgagccaaaa taataaaaaa
      241 gccggattaa taatctggct ttttatattc tctctctagt atataaacgc agaaaggccc
      301 acccgaaggt gagccagtgt gatccaagcc agctcgatat ccgtcggctt gaacgaattg
      361 ttagacatta cttgtagacg tgccctacgc gcttgttgag ttgttcctca acgtttttct
      421 tgaagtgctt agcttcaagg tcacggatac gaccgaagcg agcctcgtcc tcaatggccc
      481 gaccgattgc gcttgctaca gcctgaacgg ttgtattgtc agcactggtt aggcaagcca
      541 gagtggtctt aatggtgatg tacgctacgg cttccggctt gatttcttgc aggaactgga
      601 aggctgtcgg gcgcttgccg cgcttagctt tcacttcctc aaaccagtcg ttgatgcgtg
      661 caatcatctt agggagtagg gtagtgatga gaggcttggc ggcagcgtta tccgcaacct
      721 caccagcttt aagttgacgc tcaaacatct tgcggaagcg tgcttcaccc atctcgtaag
      781 actcatgctc aagggccaac tgttcgcgag ctaaacgctc accgtaatgg tcagccagag
      841 tgttgaacgg gatagcagcc agttcgatgt cagagaagtc gttcttagcg atgttaatcg
      901 tgttcatatt agaaaaagaa tgttattaag taagtctttc tagagctcgc tagcccaaaa
      961 aaacgggtat ggagaaacag tagagagttg cgataaaaag cgtcaggtag gatccgctaa
     1021 tcttatggat aaaaatgcta tggcatagca aagtgtgacg ccgtgcaaat aatcaatgtc
     1081 ctaggacttt tctgccgtga ttatagacac ttttgtcgcg atgttgtgtc taattttgaa
     1141 gttaactttg attccattct tttgtttgtc tgccatgatg tatacattgt gtgagttata
     1201 gttgtattcc aatttgtgtc caagaatgtt tccatcttct ttaaaatcaa taccttttaa
     1261 ctcgattcta ttaacaaggg tatcaccttc aaacttgact tcagcacgtg tcttgtagtt
     1321 cccgtcatct ttgaaaaata tagttctttc ctgtacataa ccttcgggca ctcgagacat
     1381 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     1441 ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg
     1501 ggctagcact agtggagccc tatatataaa gatcagggcc aggtatgaaa gcatttatgc
     1561 aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg tggtcttcgt
     1621 ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc attgagtcaa
     1681 ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac tctgagacta
     1741 tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg ctggctggca
     1801 tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc attactggtg
     1861 gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac agtaagaaag
     1921 cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt aacattgcgc
     1981 aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta atcaccaagt
     2041 ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc ccgatgaaac
     2101 cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct gccgctgctg
     2161 tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc atgcttgagc
     2221 aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg gactggcgcg
     2281 gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc aaaggactgc
     2341 ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg aaaatccacg
     2401 gtgcaaactg tgcgggtgtc gataaggttc cgttccctga gcgcatcaag ttcattgagg
     2461 aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact tggtgggctg
     2521 agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg gtacagcacc
     2581 acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc tctggcatcc
     2641 agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac ttgcttccta
     2701 gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag attctacaag
     2761 cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag aacactggtg
     2821 aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg ctggcttacg
     2881 gtgttactcg cagtgtgact aagcgttcag tcatgacgct ggcttacggg tccaaagagt
     2941 tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat tccggcaagg
     3001 gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg atttgggaat
     3061 ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag tctgctgcta
     3121 agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc aagcgttgcg
     3181 ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag aagcctattc
     3241 agacgcgctt ggatctgatg ttcctcggtc agttccgctt acagcctacc attaacacca
     3301 acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct aactttgtac
     3361 acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag aagtacggaa
     3421 tcgaatcttt tgcactgatt cacgactcct tcggtaccat tccggctgac gctgcgaacc
     3481 tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat gtactggctg
     3541 atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa atgccagcac
     3601 ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc gcgttcgcgt
     3661 aatctagagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt
     3721 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     3781 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     3841 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     3901 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     3961 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     4021 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     4081 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     4141 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4201 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4261 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4321 gttggggcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4381 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4441 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4501 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4561 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4621 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     4681 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4741 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4801 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4861 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4921 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4981 tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5041 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5101 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5161 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5221 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5281 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5341 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5401 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5461 agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     5521 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     5581 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg
     5641 cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca
     5701 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg
     5761 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     5821 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     5881 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     5941 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     6001 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     6061 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     6121 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     6181 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     6241 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     6301 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     6361 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     6421 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     6481 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc
     6541 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
     6601 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
     6661 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
     6721 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     6781 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     6841 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     6901 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     6961 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     7021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     7081 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     7141 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     7201 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7261 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     7321 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     7381 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     7441 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     7501 atctcaagaa gatcatctta ttaatcagat aaaatatttg ctcatgagcc cgaagtggcg
     7561 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     7621 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     7681 tc
//
