LOCUS       sequence-348388-        3540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 CDS (with stop codon) part for MoClo assembly, AATG-GCTT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3540)
  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 3540)
  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: pEPQD0CM0058
FEATURES             Location/Qualifiers
     source          1..3540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     188..206
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(210..253)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     terminator      1769..1826
                     /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(1850..1867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2021..2609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2101..2120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2792..2886)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2907..3540,1..26))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3481..3500
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
       61 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
      121 tgcccgatca actcgagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
      181 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
      241 taaggatgat ttctggaatt cggtctcaaa tggattacct taccataata ttaactttac
      301 tatttgcctt gactctctat gaagccttca gctacctatc cagaagaacc aaaaaccttc
      361 ctccaggacc atcgccattg ccgttcatcg gaagcctcca tttattaggc gaccaaccac
      421 acaaatcctt agcaaaactt tccaaaaaac acggtccaat tatgagtctc aaattaggcc
      481 agatcactac aatcgtcata tcttcatcaa caatggcgaa agaagttctt caaaaacagg
      541 atttagcatt ttcaagcaga tcagttccaa acgcactcca cgctcacaat caattcaaat
      601 tctccgttgt atggcttccg gtagcctcac gatggagaag ccttcgaaaa gttttgaatt
      661 ctaatatatt ttccggcaat cggctcgacg ctaatcaaca tttgagaact agaaaagtac
      721 aggaactaat tgcgtattgc cggaaaaata gccagagcgg agaagcggtt gacgtcggcc
      781 gagctgcttt tagaacttcg ttgaatttgt tgtcgaattt gattttttca aaggatttga
      841 cggatcctta ttcggattct gccaaggaat tcaaggattt ggtttggaat ataatggttg
      901 aggcggggaa acctaatttg gtcgattttt ttcccctgct tgaaaaagtt gatcctcaag
      961 gtatacgaca tcgtatgacg attcactttg gggaagttct taagcttttt ggtggacttg
     1021 ttaatgaaag attggagcaa agaagatcaa aaggggaaaa aaatgatgtg ttggatgtac
     1081 ttctaactac cagccaagaa agccctgagg aaatcgatag aactcacatt gagcgaatgt
     1141 gcttggacct gtttgtagca gggacggaca caacatcaag cacattagaa tgggcaatgt
     1201 cagaaatgct taaaaaccca gacaaaatga agaaaaccca agatgaactt gcacaagtaa
     1261 tcggcagagg aaaaacaata gaagaatccg atattaaccg cttaccttac ttaagatgcg
     1321 ttatgaaaga aaccttaagg atacatccac cagttccctt cttaattcct cgcaaagtgg
     1381 aacaaagtgt tgaggtttgt ggatacaatg tccctaaagg atcacaagtt cttgtgaatg
     1441 cttgggcaat tggacgtgat gaaactgttt gggatgatgc tttggcattc aaacccgaga
     1501 gatttatgga atctgaattg gatatccgtg gaagagattt cgagctgatt ccgttcggtg
     1561 ctggccgaag aatttgccca gggttgccat tggcactaag gactgtgcct ttgatgcttg
     1621 gttctttgtt gaactctttt aattggaagc ttgaaggtgg gatggctcca aaagatttgg
     1681 atatggagga gaagtttggt attacactgc agaaggctca tcctttgcgt gctgtaccaa
     1741 gcacccttta agcttcgaga ccctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct
     1801 gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg
     1861 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1921 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1981 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc
     2041 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2101 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2161 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2221 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2281 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2341 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2401 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2461 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2521 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2761 agtaaacttg gtctgacagc tcgaggcttg gattctcacc aataaaaaac gcccggcggc
     2821 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     2881 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     2941 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     3001 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     3061 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     3121 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     3181 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     3241 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     3301 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     3361 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     3421 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     3481 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
//
