LOCUS       sequence-368551-        2358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 promoter from N. benthamiana used for sgRNA expression, 
            nonstandard overlaps, GGAG-CTCG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2358)
  AUTHORS   Dudley QM, Jo S, Guerrero DAS, Chhetry M, Smedley MA, Harwood WA, 
            Sherden NH, O'Connor SE, Caputi L, Patron NJ
  TITLE     Reconstitution of monoterpene indole alkaloid biosynthesis in genome
            engineered Nicotiana benthamiana.
  JOURNAL   Commun Biol. 2022 Sep 10;5(1):949. doi: 10.1038/s42003-022-03904-w.
  PUBMED    36088516
REFERENCE   2  (bases 1 to 2358)
  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"; journalName: "Commun 
            Biol."; date: "2022-09-10"; volume: "5(1):949. doi"; pages: " 
            10.1038/s42003-022-03904-w"
COMMENT     Sequence Label: pEPQD0CM0033
FEATURES             Location/Qualifiers
     source          1..2358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..242
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(246..289)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     terminator      623..680
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(704..721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(875..1463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(955..974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1646..1740)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1761..2358,1..62))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2335..2354
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
       61 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
      121 cttatttcat tatggtgaaa gttggaacct cttacgtgcc cgatcaactc gagtgccacc
      181 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      241 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcggt
      301 ctcaggagac ataattcgtg cattcaatat atatataaat atgtatagtc aatgtatata
      361 aatggctaaa aaaataaaca ctgaatataa ctggttattt aggtaaagat ccagtcaact
      421 aatataagaa ggccggccca ttttagttaa gcccaggcac aaaaatatgg gcttattcag
      481 agggaacgaa gtgaacagta gcctagtcct gtcccaactg aaaacccaac tagctagtta
      541 gtttctccct catcggcggt gaaagaaaga ttcttcagtt tatgtagcaa accctaagat
      601 attagctcgc gagaccctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt
      661 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
      721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg
      901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      961 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1021 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1081 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1141 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1201 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1261 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1321 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1381 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1441 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1501 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1561 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1621 cttggtctga cagctcgagg cttggattct caccaataaa aaacgcccgg cggcaaccga
     1681 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1741 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     1801 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     1861 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     1921 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     1981 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     2041 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     2101 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     2161 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     2221 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     2281 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     2341 gactgaaatg cctcaaaa
//
