LOCUS       sequence-97706-e        8612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses both fragments of T7 RNAP split at position 179. Both 
            fragments are driven by the arabinose inducible promoter PBAD. Also 
            contains a constitutive araC ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8612)
  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 8612)
  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
FEATURES             Location/Qualifiers
     source          1..8612
                     /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"
     terminator      complement(1218..1245)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(4655..4672)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4825..4911
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5003..5030
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5049..5140
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5594..6049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5681..5700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(6604..7263)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7178..7197
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7264..7366)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7758..7775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7892..8437)
                     /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(7972..7991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8509..8528
                     /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 aatttcaaca cgagccaaaa taataaaaaa gccggattaa taatctggct ttttatattc
     1201 tctctctagt atataaacgc agaaaggccc acccgaaggt gagccagtgt gatccaagcc
     1261 agctcgatat ccgtcggctt gaacgaattg ttagacatta cttgtagacg tgccctacgc
     1321 gcttgttgag ttgttcctca acgtttttct tgaagtgctt agcttcaagg tcacggatac
     1381 gaccgaagcg agcctcgtcc tcaatggccc gaccgattgc gcttgctaca gcctgaacgg
     1441 ttgtattgtc agcactggtt aggcaagcca gagtggtctt aatggtgatg tacgctacgg
     1501 cttccggctt gatttcttgc aggaactgga aggctgtcgg gcgcttgccg cgcttagctt
     1561 tcacttcctc aaaccagtcg ttgatgcgtg caatcatctt agggagtagg gtagtgatga
     1621 gaggcttggc ggcagcgtta tccgcaacct caccagcttt aagttgacgc tcaaacatct
     1681 tgcggaagcg tgcttcaccc atctcgtaag actcatgctc aagggccaac tgttcgcgag
     1741 ctaaacgctc accgtaatgg tcagccagag tgttgaacgg gatagcagcc agttcgatgt
     1801 cagagaagtc gttcttagcg atgttaatcg tgttcatatt agaaaaagaa tgttattaag
     1861 taagtctttc tagagctcgc tagcccaaaa aaacgggtat ggagaaacag tagagagttg
     1921 cgataaaaag cgtcaggtag gatccgctaa tcttatggat aaaaatgcta tggcatagca
     1981 aagtgtgacg ccgtgcaaat aatcaatgtc ctaggacttt tctgccgtga ttatagacac
     2041 ttttgtcgcg atgttgtgtc taattttgaa gttaactttg attccattct tttgtttgtc
     2101 tgccatgatg tatacattgt gtgagttata gttgtattcc aatttgtgtc caagaatgtt
     2161 tccatcttct ttaaaatcaa taccttttaa ctcgattcta ttaacaaggg tatcaccttc
     2221 aaacttgact tcagcacgtg tcttgtagtt cccgtcatct ttgaaaaata tagttctttc
     2281 ctgtacataa ccttcgggca ctcgagacat tgattatttg cacggcgtca cactttgcta
     2341 tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact
     2401 ctctactgtt tctccatacc cgtttttttg ggctagcact agtggagccc tatatataaa
     2461 gatcagggcc aggtatgaaa gcatttatgc aagttgtcga ggctgacatg ctctctaagg
     2521 gtctactcgg tggcgaggcg tggtcttcgt ggcataagga agactctatt catgtaggag
     2581 tacgctgcat cgagatgctc attgagtcaa ccggaatggt tagcttacac cgccaaaatg
     2641 ctggcgtagt aggtcaagac tctgagacta tcgaactcgc acctgaatac gctgaggcta
     2701 tcgcaacccg tgcaggtgcg ctggctggca tctctccgat gttccaacct tgcgtagttc
     2761 ctcctaagcc gtggactggc attactggtg gtggctattg ggctaacggt cgtcgtcctc
     2821 tggcgctggt gcgtactcac agtaagaaag cactgatgcg ctacgaagac gtttacatgc
     2881 ctgaggtgta caaagcgatt aacattgcgc aaaacaccgc atggaaaatc aacaagaaag
     2941 tcctagcggt cgccaacgta atcaccaagt ggaagcattg tccggtcgag gacatccctg
     3001 cgattgagcg tgaagaactc ccgatgaaac cggaagacat cgacatgaat cctgaggctc
     3061 tcaccgcgtg gaaacgtgct gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc
     3121 gccgtatcag ccttgagttc atgcttgagc aagccaataa gtttgctaac cataaggcca
     3181 tctggttccc ttacaacatg gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc
     3241 cgcaaggtaa cgatatgacc aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta
     3301 aggaaggtta ctactggctg aaaatccacg gtgcaaactg tgcgggtgtc gataaggttc
     3361 cgttccctga gcgcatcaag ttcattgagg aaaaccacga gaacatcatg gcttgcgcta
     3421 agtctccact ggagaacact tggtgggctg agcaagattc tccgttctgc ttccttgcgt
     3481 tctgctttga gtacgctggg gtacagcacc acggcctgag ctataactgc tcccttccgc
     3541 tggcgtttga cgggtcttgc tctggcatcc agcacttctc cgcgatgctc cgagatgagg
     3601 taggtggtcg cgcggttaac ttgcttccta gtgaaaccgt tcaggacatc tacgggattg
     3661 ttgctaagaa agtcaacgag attctacaag cagacgcaat caatgggacc gataacgaag
     3721 tagttaccgt gaccgatgag aacactggtg aaatctctga gaaagtcaag ctgggcacta
     3781 aggcactggc tggtcaatgg ctggcttacg gtgttactcg cagtgtgact aagcgttcag
     3841 tcatgacgct ggcttacggg tccaaagagt tcggcttccg tcaacaagtg ctggaagata
     3901 ccattcagcc agctattgat tccggcaagg gtctgatgtt cactcagccg aatcaggctg
     3961 ctggatacat ggctaagctg atttgggaat ctgtgagcgt gacggtggta gctgcggttg
     4021 aagcaatgaa ctggcttaag tctgctgcta agctgctggc tgctgaggtc aaagataaga
     4081 agactggaga gattcttcgc aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc
     4141 ctgtgtggca ggaatacaag aagcctattc agacgcgctt gaacctgatg ttcctcggtc
     4201 agttccgctt acagcctacc attaacacca acaaagatag cgagattgat gcacacaaac
     4261 aggagtctgg tatcgctcct aactttgtac acagccaaga cggtagccac cttcgtaaga
     4321 ctgtagtgtg ggcacacgag aagtacggaa tcgaatcttt tgcactgatt cacgactcct
     4381 tcggtaccat tccggctgac gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg
     4441 acacatatga gtcttgtgat gtactggctg atttctacga ccagttcgct gaccagttgc
     4501 acgagtctca attggacaaa atgccagcac ttccggctaa aggtaacttg aacctccgtg
     4561 acatcttaga gtcggacttc gcgttcgcgt aatctagagt cgacctgcag gcatgcaagc
     4621 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     4681 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     4741 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     4801 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     4861 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     4921 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     4981 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     5041 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5101 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5161 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5221 gctggtgaaa gtaaaagatg ctgaagatca gttggggcaa actattaact ggcgaactac
     5281 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5341 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5401 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5461 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5521 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5581 tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5641 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5701 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5761 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5821 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5881 ttctttaata gtggactctt gttccaaact tgaacaacac tcaaccctat ctcgggctat
     5941 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6001 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta
     6061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6301 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6361 tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca cacggtcaca
     6421 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     6481 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     6541 tattatcact tattcaggcg tagcaccagg cgtttaaggg caccaataac tgccttaaaa
     6601 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     6661 gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt
     6721 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     6781 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     6841 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     6901 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     6961 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     7021 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     7081 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     7141 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     7201 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     7261 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     7321 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     7381 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     7441 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     7501 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     7561 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     7621 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     7681 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     7741 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     7801 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     7861 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     7921 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     7981 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     8041 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     8101 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     8161 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     8221 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     8281 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     8341 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     8401 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     8461 aaaatatttg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     8521 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     8581 gtagaggatc tgctcatgtt tgacagctta tc
//
