LOCUS       sequence-349012-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of truncated 3-hydroxy-3-methylglutaryl-coenzyme A 
            reductase (AstHMGR1) from Avena strigosa driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  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 6372)
  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: pEPQD1CB0817
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    131..155
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        341..686
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     629..649
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1503..1514
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2086..2260
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2305..2329
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2485..3633)
                     /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(3969..4557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4049..4068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4728..5519,5520..5588))
                     /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     5351..5370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5589..5693)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5780..6372,1..39)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gccagcccag cggcgagggc aaccagcccg gtgagcgtcg caaaggagat cctgatctga
       61 ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt
      121 ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
      181 acacattgcg gacgttttta atgtactggg gtggatgcag tgggccccac tctgtgaaga
      241 caatgccgaa ttcggatccg gaggtcaaca tggtggagca cgacactctg gtctactcca
      301 aaaatgtcaa agatacagtc tcagaagatc aaagggctat tgagactttt caacaaagga
      361 taatttcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga
      421 cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatca
      481 ttcaagatct ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
      541 tggaaaaaga agaggttcca accacgtcta caaagcaagt ggattgatgt gacatctcca
      601 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
      661 gaagttcatt tcatttggag aggacacgct cgagtataag agctcatttt tacaacaatt
      721 accaacaaca acaaacaaca aacaacatta caattacatt tacaattatc gatacaatgg
      781 cgcccgagaa aatgcccgag gaggacgagg aaatcgtcgc cggggtcgtc gcagggaaga
      841 tcccctccta cgtgctcgaa accaggctag gcgactgccg cagggcagcc gggatccgcc
      901 gcgaggcgct gcgccggatc accggcaggg agatcgacgg ccttcccctc gacggcttcg
      961 actacgactc gattctcgga cagtgctgcg agatgcccgt cgggtacgtg cagctgccgg
     1021 tcggcgtcgc ggggccgctc gtcctcgacg gccgccgcat atacgtcccg atggccacca
     1081 cggagggctg cctaatcgcc agcaccaacc gcggatgcaa ggccattgcc gagtccggag
     1141 gcgcatccag cgtcgtgtac cgcgacggga tgacccgcgc ccccgtagcc cgcttcccct
     1201 ccgcacgacg cgccgcagag ctcaagggct tcctggagaa tccggccaac tacgacaccc
     1261 tgtccgtggt ctttaacaga tcaagcagat ttgcaaggct gcagggggtc aagtgcgcca
     1321 tggctgggag gaacttgtac atgaggttca cctgcagcac cggggatgcc atggggatga
     1381 acatggtttc caagggcgtc caaaatgtgc tcgactatct gcaggaggac ttccctgaca
     1441 tggacgttgt cagcatctca ggcaactttt gttccgacaa gaaatcagct gctgtaaact
     1501 ggattgaagg ccgtggaaag tccgtggttt gtgaggcagt aatcagagag gaagttgtcc
     1561 acaaggttct caagaccaac gttcagtcac tcgtggagtt gaatgtgatc aagaaccttg
     1621 ctggctcagc agttgctggt gctcttgggg gtttcaacgc ccacgcaagc aacatcgtaa
     1681 cggctatctt cattgccact ggtcaggatc ctgcacagaa tgtggagagc tcacagtgta
     1741 tcactatgtt ggaagctgta aatgatggca gggaccttca catctccgtt acaatgccat
     1801 ctatcgaggt gggcacagtt ggtggaggca cgcagctggc ctcacagtcg gcctgcttgg
     1861 acctactggg cgtcaaaggc gccaacaggg aatctccggg gtcgaacgct aggctgctgg
     1921 ccacggtggt ggctggtgcc gtcctagctg gggagctgtc cctcatctcc gcccaagctg
     1981 ccggccatct ggtccagagc cacatgaaat acaacagatc cagcaaggac atgtccaaga
     2041 tcgcctgctg agcttctcta gctagagtcg atcgacaagc tcgagtttct ccataataat
     2101 gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct catgtgttga
     2161 gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt
     2221 ctaattccta aaaccaaaat ccagtactaa aatccagatc gctgcaattg tcttctgcac
     2281 gaagtggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     2341 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     2401 atttgtatgt gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc
     2461 agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca
     2521 ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg
     2581 ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag
     2641 cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag
     2701 cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat
     2761 ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa
     2821 gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc
     2881 cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga
     2941 ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt
     3001 gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc
     3061 gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag
     3121 cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt
     3181 tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc
     3241 cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt
     3301 tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat
     3361 ggtcatcgac ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc
     3421 ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc
     3481 cgtagcttgc tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac
     3541 ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg
     3601 cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac
     3661 tcacgccggg gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag
     3721 ataatccacc ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta
     3781 cttggtattc cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc
     3841 gtgggcctcg gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt
     3901 gtcgccggca tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg
     3961 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4021 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4081 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4141 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4201 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4261 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4321 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4381 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4441 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4501 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4561 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4621 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4681 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4741 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4801 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4861 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4921 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4981 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5041 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5101 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5161 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5221 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5281 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5341 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5401 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5461 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5521 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5581 atactcatac tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg
     5641 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat
     5701 tggccagcgc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg
     5761 gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc
     5821 ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt
     5881 ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg
     5941 caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat
     6001 ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg
     6061 cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg
     6121 aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc
     6181 ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa
     6241 gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa
     6301 cgccggcggc cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg
     6361 ccatagacgg cc
//
