LOCUS       sequence-349499-        5580 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of major latex protein+H34like (NmMLPL) from Nepeta 
            mussinii driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  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 5580)
  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: pEPQD1CB0119
FEATURES             Location/Qualifiers
     source          1..5580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(462..566)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      653..1284
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1376..1400
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        1586..1931
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1874..1894
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2539..2713
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2758..2782
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2938..4086)
                     /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(4422..5010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4502..4521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5181..5580,1..392,393..461))
                     /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"
ORIGIN
        1 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
       61 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      121 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
      181 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
      241 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
      301 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
      361 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
      421 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
      481 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag
      541 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga
      601 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc
      661 gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg
      721 cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt
      781 tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac
      841 agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc
      901 aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca
      961 cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg
     1021 ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct
     1081 gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca
     1141 gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg tctcgcacac
     1201 ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag cccagcggcg
     1261 agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat gggctgcctg
     1321 tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca
     1381 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
     1441 ttttaatgta ctggggtgga tgcagtgggc cccactctgt gaagacaatg ccgaattcgg
     1501 atccggaggt caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata
     1561 cagtctcaga agatcaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc
     1621 tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag
     1681 gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa gatctctctg
     1741 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagagg
     1801 ttccaaccac gtctacaaag caagtggatt gatgtgacat ctccactgac gtaagggatg
     1861 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     1921 tggagaggac acgctcgagt ataagagctc atttttacaa caattaccaa caacaacaaa
     1981 caacaaacaa cattacaatt acatttacaa ttatcgatac aatggcttcg aagcttgaaa
     2041 tagaaattga gttgaaaagt gatgtagaaa agatgtggaa acacttcaaa gaattcacca
     2101 aattattccc caaggcttgt cctcatcttt acgagaagat tgatgtgatc gagggagatg
     2161 ggatatcagt tggaacaatc tttgtcagta ccctgaaacc aacagagttg aaccccgtgg
     2221 ttatggtgac gaaggagaag attgattttt tagatgatga gaacaaaatg ttgaggtata
     2281 gttacatgga aggggagata ttaaagaatt acaagaattt gaggggcaca gtgcatatga
     2341 gcagcagcaa aagtggtgga actatattca aatattcagg agaattcgaa aaggcaaatg
     2401 aacaagtccc tgatcctgtt ttcttcaaag attttatggt catagttttc caaggtttgg
     2461 atgattatat tctcaagggc atgaaccata catgtcaaaa ttaagcttct ctagctagag
     2521 tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag ataagggaat
     2581 tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt
     2641 tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtac
     2701 taaaatccag atcgctgcaa ttgtcttctg cacgaagtgg tttaaactat cagtgtttga
     2761 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt
     2821 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
     2881 ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta
     2941 gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg
     3001 caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga
     3061 tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa
     3121 atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt
     3181 cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg
     3241 ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag
     3301 gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc
     3361 ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta
     3421 gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg
     3481 ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc
     3541 cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg
     3601 cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca
     3661 cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag
     3721 caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg
     3781 cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc
     3841 ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg
     3901 agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac
     3961 ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc
     4021 ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg
     4081 attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc
     4141 ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc
     4201 ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac
     4261 gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca
     4321 cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta
     4381 ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5281 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     5341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
//
