LOCUS       sequence-349418-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 4, transcriptional unit for transient 
            expression of geraniol 8-oxidase (CrG8H) from Catharanthus roseus 
            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 6756)
  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 6756)
  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: pEPQD1CB0114
FEATURES             Location/Qualifiers
     source          1..6756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    140..164
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        350..695
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     638..658
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2479..2653
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2698..2722
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2878..4026)
                     /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(4362..4950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4442..4461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5121..5912,5913..5981))
                     /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     5744..5763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5982..6086)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6173..6756,1..48)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tagacggccg ccagcccagc ggcgagggca accagcccgg tgagcgtcgc aaaggagatc
       61 ctgatctgac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg
      121 ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac
      181 aacttaataa cacattgcgg acgtttttaa tgtactgggg tggatgcagt gggccccact
      241 ctgtgaagac aattacgaat tcccatgggg aggtcaacat ggtggagcac gacactctgg
      301 tctactccaa aaatgtcaaa gatacagtct cagaagatca aagggctatt gagacttttc
      361 aacaaaggat aatttcggga aacctcctcg gattccattg cccagctatc tgtcacttca
      421 tcgaaaggac agtagaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa
      481 aggctatcat tcaagatctc tctgccgaca gtggtcccaa agatggaccc ccacccacga
      541 ggagcatcgt ggaaaaagaa gaggttccaa ccacgtctac aaagcaagtg gattgatgtg
      601 acatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct
      661 ctatataagg aagttcattt catttggaga ggacacgcta ctgtataaga gctcattttt
      721 acaacaatta ccaacaacaa caaacaacaa acaacattac aattacattt acaattatcg
      781 ataacatggc ttcttctatg ctttcttctg ctgctgttgt tgctactcgt gctagtgctg
      841 ctcaagctag tatggttgct ccttttactg gacttaagtc tgctgcttct tttcctgtta
      901 ctagaaagca aaacaacctt gatattactt ctattgctag taacggagga agagttcgag
      961 caatggatta ccttaccata atattaactt tactatttgc cttgactctc tatgaagcct
     1021 tcagctacct atccagaaga accaaaaacc ttcctccagg accatcgcca ttgccgttca
     1081 tcggaagcct ccatttatta ggcgaccaac cacacaaatc cttagcaaaa ctttccaaaa
     1141 aacacggtcc aattatgagt ctcaaattag gccagatcac tacaatcgtc atatcttcat
     1201 caacaatggc gaaagaagtt cttcaaaaac aggatttagc attttcaagc agatcagttc
     1261 caaacgcact ccacgctcac aatcaattca aattctccgt tgtatggctt ccggtagcct
     1321 cacgatggag aagccttcga aaagttttga attctaatat attttccggc aatcggctcg
     1381 acgctaatca acatttgaga actagaaaag tacaggaact aattgcgtat tgccggaaaa
     1441 atagccagag cggagaagcg gttgacgtcg gccgagctgc ttttagaact tcgttgaatt
     1501 tgttgtcgaa tttgattttt tcaaaggatt tgacggatcc ttattcggat tctgccaagg
     1561 aattcaagga tttggtttgg aatataatgg ttgaggcggg gaaacctaat ttggtcgatt
     1621 tttttcccct gcttgaaaaa gttgatcctc aaggtatacg acatcgtatg acgattcact
     1681 ttggggaagt tcttaagctt tttggtggac ttgttaatga aagattggag caaagaagat
     1741 caaaagggga aaaaaatgat gtgttggatg tacttctaac taccagccaa gaaagccctg
     1801 aggaaatcga tagaactcac attgagcgaa tgtgcttgga cctgtttgta gcagggacgg
     1861 acacaacatc aagcacatta gaatgggcaa tgtcagaaat gcttaaaaac ccagacaaaa
     1921 tgaagaaaac ccaagatgaa cttgcacaag taatcggcag aggaaaaaca atagaagaat
     1981 ccgatattaa ccgcttacct tacttaagat gcgttatgaa agaaacctta aggatacatc
     2041 caccagttcc cttcttaatt cctcgcaaag tggaacaaag tgttgaggtt tgtggataca
     2101 atgtccctaa aggatcacaa gttcttgtga atgcttgggc aattggacgt gatgaaactg
     2161 tttgggatga tgctttggca ttcaaacccg agagatttat ggaatctgaa ttggatatcc
     2221 gtggaagaga tttcgagctg attccgttcg gtgctggccg aagaatttgc ccagggttgc
     2281 cattggcact aaggactgtg cctttgatgc ttggttcttt gttgaactct tttaattgga
     2341 agcttgaagg tgggatggct ccaaaagatt tggatatgga ggagaagttt ggtattacac
     2401 tgcagaaggc tcatcctttg cgtgctgtac caagcaccct ttaagcttct ctagctagag
     2461 tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag ataagggaat
     2521 tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt
     2581 tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtac
     2641 taaaatccag atcgctcaga ttgtcttctg cacgaagtgg tttaaactat cagtgtttga
     2701 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt
     2761 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
     2821 ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta
     2881 gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg
     2941 caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga
     3001 tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa
     3061 atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt
     3121 cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg
     3181 ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag
     3241 gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc
     3301 ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta
     3361 gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg
     3421 ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc
     3481 cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg
     3541 cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca
     3601 cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag
     3661 caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg
     3721 cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc
     3781 ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg
     3841 agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac
     3901 ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc
     3961 ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg
     4021 attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc
     4081 ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc
     4141 ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac
     4201 gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca
     4261 cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta
     4321 ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4921 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5041 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5101 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5161 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5221 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     5281 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5341 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5401 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5461 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5521 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5581 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5641 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5701 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5761 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5821 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5881 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5941 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6001 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag
     6061 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga
     6121 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc
     6181 gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg
     6241 cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt
     6301 tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac
     6361 agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc
     6421 aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca
     6481 cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg
     6541 ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct
     6601 gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca
     6661 gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg tctcgcacac
     6721 ggcttcgacg gcgtttctgg cgcgtttgca gggcca
//
