LOCUS       sequence-349500-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of major latex protein-like (NmMLPL) from Nepeta mussinii
            driven by 35S promoter; contains a chloroplast transit peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  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 5757)
  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: pEPQD1CB0120
FEATURES             Location/Qualifiers
     source          1..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    158..182
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        368..713
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     656..676
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1498..1672
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    1717..1741
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1897..3045)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3381..3969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3461..3480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4140..4931,4932..5000))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4763..4782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5001..5105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5192..5757,1..66)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ggcgcgtttg cagggccata gacggccgcc agcccagcgg cgagggcaac cagcccggtg
       61 agcgtcgcaa aggagatcct gatctgactg atgggctgcc tgtatcgagt ggtgattttg
      121 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa
      181 acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg tactggggtg
      241 gatgcagtgg gccccactct gtgaagacaa tgccgaattc ggatccggag gtcaacatgg
      301 tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa
      361 gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc
      421 cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc
      481 atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag
      541 atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa
      601 agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa tcccactatc
      661 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acacgctact
      721 gtataagagc tcatttttac aacaattacc aacaacaaca aacaacaaac aacattacaa
      781 ttacatttac aattatcgat aacatggctt cttctatgct ttcttctgct gctgttgttg
      841 ctactcgtgc tagtgctgct caagctagta tggttgctcc ttttactgga cttaagtctg
      901 ctgcttcttt tcctgttact agaaagcaaa acaaccttga tattacttct attgctagta
      961 acggaggaag agttcgagca atggcttcga agcttgaaat agaaattgag ttgaaaagtg
     1021 atgtagaaaa gatgtggaaa cacttcaaag aattcaccaa attattcccc aaggcttgtc
     1081 ctcatcttta cgagaagatt gatgtgatcg agggagatgg gatatcagtt ggaacaatct
     1141 ttgtcagtac cctgaaacca acagagttga accccgtggt tatggtgacg aaggagaaga
     1201 ttgatttttt agatgatgag aacaaaatgt tgaggtatag ttacatggaa ggggagatat
     1261 taaagaatta caagaatttg aggggcacag tgcatatgag cagcagcaaa agtggtggaa
     1321 ctatattcaa atattcagga gaattcgaaa aggcaaatga acaagtccct gatcctgttt
     1381 tcttcaaaga ttttatggtc atagttttcc aaggtttgga tgattatatt ctcaagggca
     1441 tgaaccatac atgtcaaaat taagcttctc tagctagagt cgatcgacaa gctcgagttt
     1501 ctccataata atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg
     1561 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     1621 tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tcgctgcaat
     1681 tgtcttctgc acgaagtggt ttaaactatc agtgtttgac aggatatatt ggcgggtaaa
     1741 cctaagagaa aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt
     1801 atccgttcgt ccatttgtat gtgcatgcca accacagggt tccccagatc aggcgctggc
     1861 tgctgaaccc ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc
     1921 atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc
     1981 gacctgctcg cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt
     2041 ttccagcttg agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc
     2101 cgtcggcgac agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa
     2161 cagcacgacg atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc
     2221 caggacgcgg aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt
     2281 gaagcccatc gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg
     2341 gccattgatc gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc
     2401 gccgataggg gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc
     2461 ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac
     2521 cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg
     2581 ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga
     2641 aagctgcatt tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcctc
     2701 gctgacctgt tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc
     2761 tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc tcctgttcaa gacgacgcga
     2821 acgctccacg gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg
     2881 ctcgatcttg gccgtagctt gctggaccat cgagccgacg gactggaagg tttcgcgggg
     2941 cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc
     3001 gatcagttct tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg
     3061 gattgccccg actcacgccg gggcaatgtg cccttattcc tgatttgacc cgcctggtgc
     3121 cttggtgtcc agataatcca ccttatcggc aatgaagtcg gtcccgtaga ccgtctggcc
     3181 gtccttctcg tacttggtat tccgaatctt gccctgcacg aataccagcg accccttgcc
     3241 caaatacttg ccgtgggcct cggcctgaga gccaaaacac ttgatgcgga agaagtcggt
     3301 gcgctcctgc ttgtcgccgg catcgttgcg ccacatctag gatctgccag gaaccgtaaa
     3361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3781 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4261 agtgctgcaa tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac
     4321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcaaggt
     5041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5101 ccgcgcacga attggccagc gctgccattt ttggggtgag gccgttcgcg gccgaggggc
     5161 gcagcccctg gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg
     5221 gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg
     5281 tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg
     5341 cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg
     5401 tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg
     5461 tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac
     5521 atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag
     5581 ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag
     5641 tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg
     5701 aggtatccac aacgccggcg gccgcggtgt ctcgcacacg gcttcgacgg cgtttct
//
