LOCUS       sequence-97707-e        6040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the N-terminal fragment of split T7 RNAP split a position 
            179. Insert is driven by PBAD. Contains a constitutive araC ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6040)
  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 6040)
  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-N
FEATURES             Location/Qualifiers
     source          1..6040
                     /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"
     terminator      1939..1966
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2253..2339
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2431..2458
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2477..2568
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3022..3477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3109..3128)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(4032..4691)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4606..4625
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4692..4794)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5186..5203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5320..5865)
                     /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(5400..5419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5937..5956
                     /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 ctagcgagct ctagaaagac
     1321 ttacttaata acattctttt tctaatatga acacgattaa catcgctaag aacgacttct
     1381 ctgacatcga actggctgct atcccgttca acactctggc tgaccattac ggtgagcgtt
     1441 tagctcgcga acagttggcc cttgagcatg agtcttacga gatgggtgaa gcacgcttcc
     1501 gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc ggataacgct gccgccaagc
     1561 ctctcatcac taccctactc cctaagatga ttgcacgcat caacgactgg tttgaggaag
     1621 tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt cctgcaagaa atcaagccgg
     1681 aagccgtagc gtacatcacc attaagacca ctctggcttg cctaaccagt gctgacaata
     1741 caaccgttca ggctgtagca agcgcaatcg gtcgggccat tgaggacgag gctcgcttcg
     1801 gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa cgttgaggaa caactcaaca
     1861 agcgcgtagg gcacgtctac aagtaatgtc taacaattcg ttcaagccga cggatatcga
     1921 gctggcttgg atcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag
     1981 agagaatata aaaagccaga ttattaatcc ggctttttta ttattttggc tcgtgttgaa
     2041 atcgcgatgt tgtgtctaat tttgaagtta actttgattc cattcttttg tttgtctgcc
     2101 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     2161 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     2221 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     2281 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2341 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     2401 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
     2461 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tggggcaaac tattaactgg
     2701 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2761 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2821 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2881 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2941 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3001 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3061 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3121 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3181 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3241 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3301 agtccacgtt ctttaatagt ggactcttgt tccaaacttg aacaacactc aaccctatct
     3361 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3421 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     3481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3721 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tcaggcattt gagaagcaca
     3841 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     3901 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
     3961 catccgctta ttatcactta ttcaggcgta gcaccaggcg tttaagggca ccaataactg
     4021 ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     4081 attctgccga catggaagcc atcacagacg gcatgatgaa cctgaatcgc cagcggcatc
     4141 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     4201 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     4261 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     4321 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     4381 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     4441 atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg
     4501 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     4561 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     4621 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     4681 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     4741 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct
     4801 cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt
     4861 tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat
     4921 gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag tggcttctgt
     4981 ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac
     5041 cgccggacat cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt
     5101 gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata
     5161 tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
     5221 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
     5281 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
     5341 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg
     5461 gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca
     5521 gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg
     5581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag
     5641 caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc
     5701 ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct
     5761 cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt
     5821 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt
     5881 aatcagataa aatatttgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5941 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     6001 cgtccggcgt agaggatctg ctcatgtttg acagcttatc
//
