LOCUS       sequence-349503-        6546 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MoClo Level 1, position 3, transcriptional unit for transient 
            expression of 7-deoxyloganetic acid glucosyl transferase (Cr7-DLGT) 
            from Catharanthus roseus driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6546)
  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 6546)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pEPQD1CB0123
FEATURES             Location/Qualifiers
     source          1..6546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    167..191
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        377..722
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     665..685
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2296..2470
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2515..2539
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2695..3843)
                     /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(4179..4767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4259..4278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4938..5729,5730..5798))
                     /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     5561..5580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5799..5903)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5990..6546,1..75)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ggcgtttctg gcgcgtttgc agggccatag acggccgcca gcccagcggc gagggcaacc
       61 agcccggtga gcgtcgcaaa ggagatcctg atctgactga tgggctgcct gtatcgagtg
      121 gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt
      181 gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg tttttaatgt
      241 actggggtgg atgcagtggg ccccactctg tgaagacaaa ctagaattcg agctcggagg
      301 tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag
      361 aagatcaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat
      421 tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct
      481 acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatctctct gccgacagtg
      541 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagag gttccaacca
      601 cgtctacaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat
      661 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
      721 cacgctcgag tataagagct catttttaca acaattacca acaacaacaa acaacaaaca
      781 acattacaat tacatttaca attatcgata caatgggttc tcaagaaaca aatttggctc
      841 ctcatgttct catattccct ttgccaattc aaggccatgt gaactccatg ctcagactag
      901 cagaacttct ctgtctagct gaattagata tcacttttat tgtttcagag ttcagccaca
      961 gtcgcttaat taagcataca aatgttgcct cccgttttgc ccgttaccca ggatttcagt
     1021 tccagcccat atccgatggc cttcccgatg atcaccccag agcaggcgaa agagtcatgg
     1081 atattttgcc ttcaactaaa aatgtaacgg gacccctgtt taagcagatg atggttgaga
     1141 ataagtgctt ttcttctgct actcgaaggc cgattacttg cataatcgct gatggggttt
     1201 tgagctttgc tggggatttc gctcaagaaa aaggaatccc tcttatttac ttcagaactg
     1261 tttctgcttg ttctttctgg gcttgttttt gtatgcctga actaattgaa tctggtgaca
     1321 ttccgattaa aggaaatggt atggatctga tagtgaaaag cgtgccgggg atggaaactt
     1381 ttctcaggcg acgagatctg ccggggtttt gccgagtcaa tgacattaat gaaccgaaac
     1441 tccagatcct gaaaacagag acaagacaga caacgagagc ccaagcggct attttgaaca
     1501 cttttgagga tttggaagga ccaattctat ctcaaattcg caaacacatg ccaagacttt
     1561 tcacaattgg accaagtcat tcccacctaa catctagact tgaaaccaag aatatcaaga
     1621 ctttaatttc ctccggtagt ttctgggaag aagatcgaag ctgtgtggat tggcttgatg
     1681 cacagccacc aagatctgtt ttatatgtaa gttttggaag tattacagtt gtaacaagag
     1741 atcaactcct ggaattctgg tatggtcttg ttaatagtgg acaacggttc ttgtgggtaa
     1801 tgaggccaga ttcaattatg ggcaaagatg ggcaaagcca gattcctgca gatcttgaag
     1861 agggcacaaa agccagaggt tatatggtgg gatgggcacc tcaagaagag gtcttgaatc
     1921 accctgccat tggaggattt ctgactcaca gtgggtggaa ttcaacttta gagagtattg
     1981 tggctggtgt cccaatgata tgttggccat actttgcgga ccaaatgata aacagcagat
     2041 ttgtgagtga aatttggaag attggattgg atatgaaaga tacatgtgat agagagacaa
     2101 ttgtgaagat ggttagggag ttgatggaga ttaggaaaga tgagttcttg caaagggctg
     2161 atcatatggc taaattggca aaagaggctg ttagtgaagg cggatcctca tattctaact
     2221 tggatggcct cgttgattac ataaaatcac tgattatttg agcttctcta gctagagtcg
     2281 atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag
     2341 ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt
     2401 atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa
     2461 aatccagatc gctttacttg tcttctgcac gaagtggttt aaactatcag tgtttgacag
     2521 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     2581 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     2641 cccagatcag gcgctggctg ctgaaccccc agccggaact gaccccacaa ggccctagcg
     2701 tttgcaatgc accaggtcat cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa
     2761 ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc
     2821 gcacatgagg cggaaggttt ccagcttgag cgggtacggc tcccggtgcg agctgaaata
     2881 gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac ttctcccata tgaatttcgt
     2941 gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct ggcaacggga
     3001 cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg attccaggtg
     3061 cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc
     3121 ggccttcgtg taataccggc cattgatcga ccagcccagg tcctggcaaa gctcgtagaa
     3181 cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg tactccaaca cctgctgcca
     3241 caccagttcg tcatcgtcgg cccgcagctc gacgccggtg taggtgatct tcacgtcctt
     3301 gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt
     3361 ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg
     3421 cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa
     3481 cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt
     3541 cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc
     3601 ctgttcaaga cgacgcgaac gctccacggc ggccgatggc gcgggcaggg cagggggagc
     3661 cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc tggaccatcg agccgacgga
     3721 ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt cggcatcctc
     3781 ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt
     3841 cattcaccct ccttgcggga ttgccccgac tcacgccggg gcaatgtgcc cttattcctg
     3901 atttgacccg cctggtgcct tggtgtccag ataatccacc ttatcggcaa tgaagtcggt
     3961 cccgtagacc gtctggccgt ccttctcgta cttggtattc cgaatcttgc cctgcacgaa
     4021 taccagcgac cccttgccca aatacttgcc gtgggcctcg gcctgagagc caaaacactt
     4081 gatgcggaag aagtcggtgc gctcctgctt gtcgccggca tcgttgcgcc acatctagga
     4141 tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4201 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4261 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4321 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4381 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4441 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4501 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4561 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4621 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4681 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4741 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4801 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4861 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4921 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4981 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5041 ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc accggctcca
     5101 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5161 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5221 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5281 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5341 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5401 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5461 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5521 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5581 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5641 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5701 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5761 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5821 tgaagcattt atcaaggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5881 aataaacaaa taggggttcc gcgcacgaat tggccagcgc tgccattttt ggggtgaggc
     5941 cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta gcgggccggg
     6001 agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca cagggcgcag
     6061 ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa gacaggttag
     6121 cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc tgtggacagc
     6181 ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc aagtgtcaag
     6241 gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg cagggcactt
     6301 atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc gttttcgccg
     6361 atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc ctcatctgtc
     6421 aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca tctgtcagtg
     6481 agggccaagt tttccgcgag gtatccacaa cgccggcggc cgcggtgtct cgcacacggc
     6541 ttcgac
//
