LOCUS       sequence-227369-        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"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      2593..2624
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2640..3299)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      complement(3452..4190)
                     /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      4280..4325
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4340..5218)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac ccaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttgga
      301 cgcgtacaac tcaagtctga cataaatgac cgctatggca agcgagcagc tggaaaagaa
      361 gttacaagcc ctggagaaaa aacttgctca gctggaatgg aaaaaccaag cattggaaaa
      421 aaaactcgcg cagacgtcgg gtggaagcgg taaagcattt atgcaagttg tcgaggctga
      481 catgctctct aagggtctac tcggtggcga ggcgtggtct tcgtggcata aggaagactc
      541 tattcatgta ggagtacgct gcatcgagat gctcattgag tcaaccggaa tggttagctt
      601 acaccgccaa aatgctggcg tagtaggtca agactctgag actatcgaac tcgcacctga
      661 atacgctgag gctatcgcaa cccgtgcagg tgcgctggct ggcatctctc cgatgttcca
      721 accttgcgta gttcctccta agccgtggac tggcattact ggtggtggct attgggctaa
      781 cggtcgtcgt cctctggcgc tggtgcgtac tcacagtaag aaagcactga tgcgctacga
      841 agacgtttac atgcctgagg tgtacaaagc gattaacatt gcgcaaaaca ccgcatggaa
      901 aatcaacaag aaagtcctag cggtcgccaa cgtaatcacc aagtggaagc attgtccggt
      961 cgaggacatc cctgcgattg agcgtgaaga actcccgatg aaaccggaag acatcgacat
     1021 gaatcctgag gctctcaccg cgtggaaacg tgctgccgct gctgtgtacc gcaaggacaa
     1081 ggctcgcaag tctcgccgta tcagccttga gttcatgctt gagcaagcca ataagtttgc
     1141 taaccataag gccatctggt tcccttacaa catggactgg cgcggtcgtg tttacgctgt
     1201 gtcaatgttc aacccgcaag gtaacgatat gaccaaagga ctgcttacgc tggcgaaagg
     1261 taaaccaatc ggtaaggaag gttactactg gctgaaaatc cacggtgcaa actgtgcggg
     1321 tgtcgataag gttccgttcc ctgagcgcat caagttcatt gaggaaaacc acgagaacat
     1381 catggcttgc gctaagtctc cactggagaa cacttggtgg gctgagcaag attctccgtt
     1441 ctgcttcctt gcgttctgct ttgagtacgc tggggtacag caccacggcc tgagctataa
     1501 ctgctccctt ccgctggcgt ttgacgggtc ttgctctggc atccagcact tctccgcgat
     1561 gctccgagat gaggtaggtg gtcgcgcggt taacttgctt cctagtgaaa ccgttcagga
     1621 catctacggg attgttgcta agaaagtcaa cgagattcta caagcagacg caatcaatgg
     1681 gaccgataac gaagtagtta ccgtgaccga tgagaacact ggtgaaatct ctgagaaagt
     1741 caagctgggc actaaggcac tggctggtca atggctggct tacggtgtta ctcgcagtgt
     1801 gactaagcgt tcagtcatga cgctggctta cgggtccaaa gagttcggct tccgtcaaca
     1861 agtgctggaa gataccattc agccagctat tgattccggc aagggtctga tgttcactca
     1921 gccgaatcag gctgctggat acatggctaa gctgatttgg gaatctgtga gcgtgacggt
     1981 ggtagctgcg gttgaagcaa tgaactggct taagtctgct gctaagctgc tggctgctga
     2041 ggtcaaagat aagaagactg gagagattct tcgcaagcgt tgcgctgtgc attgggtaac
     2101 tcctgatggt ttccctgtgt ggcaggaata caagaagcct attcagacgc gcttgaacct
     2161 gatgttcctc ggtcagttcc gcttacagcc taccattaac accaacaaag atagcgagat
     2221 tgatgcacac aaacaggagt ctggtatcgc tcctaacttt gtacacagcc aagacggtag
     2281 ccaccttcgt aagactgtag tgtgggcaca cgagaagtac ggaatcgaat cttttgcact
     2341 gattcacgac tccttcggta ccattccggc tgacgctgcg aacctgttca aagcagtgcg
     2401 cgaaactatg gttgacacat atgagtcttg tgatgtactg gctgatttct acgaccagtt
     2461 cgctgaccag ttgcacgagt ctcaattgga caaaatgcca gcacttccgg ctaaaggtaa
     2521 cttgaacctc cgtgacatct tagagtcgga cttcgcgttc gcgtaaactt aattaacggc
     2581 actcctcagc caagtcaaaa gcctccgacc ggaggctttt gactacatgc ccatggcgtt
     2641 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     2701 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     2761 ccttgcgtat aatatttgcc catagtgaaa acgggggcga agaagttgtc catattggcc
     2821 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     2881 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     2941 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     3001 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     3061 ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc aagaatgtga
     3121 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     3181 tccagctgaa cggtctggtt ataggtacat tgagtaactg actgaaatgc ctcaaaatgt
     3241 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     3301 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     3361 atttcattat ggtgaaagtt ggaacctctt acgtgccaag ccaaataggc cgtcactcgg
     3421 tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac
     3481 agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg
     3541 gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt
     3601 gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag
     3661 gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac
     3721 cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct
     3781 catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta
     3841 agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt
     3901 ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg
     3961 cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta
     4021 cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag
     4081 ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag
     4141 ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg
     4201 aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccag
     4261 gaggaagagg acatccggtc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4321 ttttagactt agggaccctt tatgacaact tgacggctac atcattcact ttttcttcac
     4381 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     4441 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     4501 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     4561 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     4621 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     4681 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     4741 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
     4801 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     4861 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     4921 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     4981 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     5041 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     5101 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     5161 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     5221 ttttcatact cccaccattc agag
//
