LOCUS       sequence-222991-        5244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZB-linker-split C-terminal T7 RNAP expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5244)
  AUTHORS   Pu J, Zinkus-Boltz J, Dickinson BC
  TITLE     Evolution of a split RNA polymerase as a versatile biosensor 
            platform.
  JOURNAL   Nat Chem Biol. 2017 Apr;13(4):432-438. doi: 10.1038/nchembio.2299. 
            Epub 2017 Feb 13.
  PUBMED    28192413
REFERENCE   2  (bases 1 to 5244)
  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: 
            "10.1038/nchembio.2299"; journalName: "Nat Chem Biol"; date: 
            "2017-04"; volume: "13"; issue: "4"; pages: "432-438"
COMMENT     Sequence Label: p2-39
FEATURES             Location/Qualifiers
     source          1..5244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..777)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     terminator      867..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(927..1805)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1832..2116
                     /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     2039..2058
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      4424..4455
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4471..5130)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
       61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
      121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
      181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
      241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
      301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
      361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
      421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
      481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
      541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
      601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
      661 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
      721 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
      841 agcccaggag gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tagacttagg gaccctttat gacaacttga cggctacatc attcactttt
      961 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     1021 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     1081 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     1141 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     1201 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     1261 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     1321 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     1381 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     1441 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     1501 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     1561 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     1621 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     1681 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     1741 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     1801 gccatacttt tcatactccc accattcaga gaagaaacca attgtccata ttgcatcaga
     1861 cattgccgtc actgcgtctt ttactggctc ttctcgctaa cccaaccggt aaccccgctt
     1921 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     1981 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
     2041 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
     2101 tctactgttt ctccataccc gtttttttgg acgcgtacaa ctcaagtctg acataaatga
     2161 ccgctatggc aagcgagcag ctggaaaaga agttacaagc cctggagaaa aaacttgctc
     2221 agctggaatg gaaaaaccaa gcattggaaa aaaaactcgc gcagacgtcg ggtggaagcg
     2281 gtaaagcatt tatgcaagtt gtcgaggctg acatgctctc taagggtcta ctcggtggcg
     2341 aggcgtggtc ttcgtggcat aaggaagact ctattcatgt aggagtacgc tgcatcgaga
     2401 tgctcattga gtcaaccgga atggttagct tacaccgcca aaatgctggc gtagtaggtc
     2461 aagactctga gactatcgaa ctcgcacctg aatacgctga ggctatcgca acccgtgcag
     2521 gtgcgctggc tggcatctct ccgatgttcc aaccttgcgt agttcctcct aagccgtgga
     2581 ctggcattac tggtggtggc tattgggcta acggtcgtcg tcctctggcg ctggtgcgta
     2641 ctcacagtaa gaaagcactg atgcgctacg aagacgttta catgcctgag gtgtacaaag
     2701 cgattaacat tgcgcaaaac accgcatgga aaatcaacaa gaaagtccta gcggtcgcca
     2761 acgtaatcac caagtggaag cattgtccgg tcgaggacat ccctgcgatt gagcgtgaag
     2821 aactcccgat gaaaccggaa gacatcgaca tgaatcctga ggctctcacc gcgtggaaac
     2881 gtgctgccgc tgctgtgtac cgcaaggaca aggctcgcaa gtctcgccgt atcagccttg
     2941 agttcatgct tgagcaagcc aataagtttg ctaaccataa ggccatctgg ttcccttaca
     3001 acatggactg gcgcggtcgt gtttacgctg tgtcaatgtt caacccgcaa ggtaacgata
     3061 tgaccaaagg actgcttacg ctggcgaaag gtaaaccaat cggtaaggaa ggttactact
     3121 ggctgaaaat ccacggtgca aactgtgcgg gtgtcgataa ggttccgttc cctgagcgca
     3181 tcaagttcat tgaggaaaac cacgagaaca tcatggcttg cgctaagtct ccactggaga
     3241 acacttggtg ggctgagcaa gattctccgt tctgcttcct tgcgttctgc tttgagtacg
     3301 ctggggtaca gcaccacggc ctgagctata actgctccct tccgctggcg tttgacgggt
     3361 cttgctctgg catccagcac ttctccgcga tgctccgaga tgaggtaggt ggtcgcgcgg
     3421 ttaacttgct tcctagtgaa accgttcagg acatctacgg gattgttgct aagaaagtca
     3481 acgagattct acaagcagac gcaatcaatg ggaccgataa cgaagtagtt accgtgaccg
     3541 atgagaacac tggtgaaatc tctgagaaag tcaagctggg cactaaggca ctggctggtc
     3601 aatggctggc ttacggtgtt actcgcagtg tgactaagcg ttcagtcatg acgctggctt
     3661 acgggtccaa agagttcggc ttccgtcaac aagtgctgga agataccatt cagccagcta
     3721 ttgattccgg caagggtctg atgttcactc agccgaatca ggctgctgga tacatggcta
     3781 agctgatttg ggaatctgtg agcgtgacgg tggtagctgc ggttgaagca atgaactggc
     3841 ttaagtctgc tgctaagctg ctggctgctg aggtcaaaga taagaagact ggagagattc
     3901 ttcgcaagcg ttgcgctgtg cattgggtaa ctcctgatgg tttccctgtg tggcaggaat
     3961 acaagaagcc tattcagacg cgcttgaacc tgatgttcct cggtcagttc cgcttacagc
     4021 ctaccattaa caccaacaaa gatagcgaga ttgatgcaca caaacaggag tctggtatcg
     4081 ctcctaactt tgtacacagc caagacggta gccaccttcg taagactgta gtgtgggcac
     4141 acgagaagta cggaatcgaa tcttttgcac tgattcacga ctccttcggt accattccgg
     4201 ctgacgctgc gaacctgttc aaagcagtgc gcgaaactat ggttgacaca tatgagtctt
     4261 gtgatgtact ggctgatttc tacgaccagt tcgctgacca gttgcacgag tctcaattgg
     4321 acaaaatgcc agcacttccg gctaaaggta acttgaacct ccgtgacatc ttagagtcgg
     4381 acttcgcgtt cgcgtaaact taattaacgg cactcctcag ccaagtcaaa agcctccgac
     4441 cggaggcttt tgactacatg cccatggcgt ttacgccccg ccctgccact catcgcagta
     4501 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     4561 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa
     4621 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     4681 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     4741 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     4801 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     4861 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg
     4921 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     4981 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     5041 ttgagtaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     5101 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     5161 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     5221 tacgtgccaa gccaaatagg ccgt
//
