LOCUS       sequence-349006-        2303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 promoter binding site part for MoClo assembly, nonstandard 
            overlaps, CCAT-TACT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2303)
  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 2303)
  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: pEPKK0CM0480
FEATURES             Location/Qualifiers
     source          1..2303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..160
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     388..406
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(410..453)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    485..518
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    544..577
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        594..640
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      732..789
                     /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(813..830)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(984..1572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1064..1083)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1755..1849)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1870..2303,1..226))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
       61 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
      121 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
      181 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
      241 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
      301 gaaagttgga acctcttacg tgcccgatca actcgagtgc cacctgacgt ctaagaaacc
      361 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa
      421 aaaaaatcct tagctttcgc taaggatgat ttctggaatt cggtctcacc atggtgcggt
      481 gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta ctccactccc cgagccggtg
      541 cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccactcccc ttcgcaagac
      601 ccttcctcta tataaggaag ttcatttcat ttggagagga ctccggtatt tttacaacaa
      661 ttaccacaac aaaacaaaca acaaacaaca ttacaattta ctattctagt cgaatactcg
      721 agaccctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa
      781 ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc
      841 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      961 aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa
     1021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1381 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1681 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1741 agctcgaggc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     1801 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     1861 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     1921 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     1981 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     2041 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     2101 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     2161 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     2221 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     2281 caccagctca ccgtctttca ttg
//
