LOCUS       sequence-349018-        6228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 2, transcriptional unit for transient 
            expression of geranyl pyrophosphate synthase (PaGPPS1) from Picea 
            abies 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 6228)
  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 6228)
  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: pEPQD1CB0109
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    123..147
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        333..678
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     621..641
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1934..2108
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2153..2177
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2333..3481)
                     /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(3817..4405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3897..3916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4576..5367,5368..5436))
                     /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     5199..5218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5437..5541)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5628..6228,1..31)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcctgatct gactgatggg
       61 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
      121 ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacacattg
      181 cggacgtttt taatgtactg gggtggatgc agtgggcccc actctgtgaa gacaagcaag
      241 aattcaagct tggaggtcaa catggtggag cacgacactc tggtctactc caaaaatgtc
      301 aaagatacag tctcagaaga tcaaagggct attgagactt ttcaacaaag gataatttcg
      361 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa
      421 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat
      481 ctctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
      541 gaagaggttc caaccacgtc tacaaagcaa gtggattgat gtgacatctc cactgacgta
      601 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
      661 tttcatttgg agaggacacg ctactgtata agagctcatt tttacaacaa ttaccaacaa
      721 caacaaacaa caaacaacat tacaattaca tttacaatta tcgataacat ggcttcttct
      781 atgctttctt ctgctgctgt tgttgctact cgtgctagtg ctgctcaagc tagtatggtt
      841 gctcctttta ctggacttaa gtctgctgct tcttttcctg ttactagaaa gcaaaacaac
      901 cttgatatta cttctattgc tagtaacgga ggaagagttc gagcaatgtg ctcaaacaca
      961 aatgcccagt tggccaattt gtctgaacaa gtaaagggga aggtcactga atttgatttc
     1021 aaggagtaca tgcgttccaa ggcaatgtca gtgaatgagg cactggacag ggctgtccca
     1081 cttcgttatc ctgaaaaaat acatgaagct atgaggtatt ctcttctagc aggagggaag
     1141 cgagttcgtc ctattctgtg cattgcagca tgtgagcttg tgggagggag cgaggagctc
     1201 gccatgccaa ctgcctgtgc aatggaaatt atccacacaa tgtctttgat tcatgatgac
     1261 ttgcctccca tggacaatga tgatttacgc cgaggtaagc ctacaaacca caaggtgttc
     1321 ggtgaaggca ctgctgttct tgcgggggat gcacttcttt catttgcatt tgagcacatt
     1381 gcagtgtcca caagcaagac tgtggagtct gatagggttt tgagggtggt atctgaattg
     1441 ggtagagcaa taggctcgga aggggttgca ggtgggcagg ttgctgatat taccagtcag
     1501 gggaatcctt ctgttggcct tgagactctg gaatggattc acattcacaa gactgcagtg
     1561 ctcttggagt gctccgttgc tagtggggcg atcatcggtg gtgcttctga tgatgagatt
     1621 gagagagttc gaaagtatgc ccgttgcgtg gggcttctgt ttcaggttgt ggatgacatt
     1681 ctcgatgtca cgaaatcatc agaagaattg ggtaagactg cagcaaagga tttgcttagt
     1741 gataaggcca cttatcccaa gctcatgggt ttggagaaag caaaggagtt tgctgatgaa
     1801 ttgttgggca aagctaagga ggagttatct ttcttcaacc caactaaggc tgcacctctg
     1861 ttgggtcttg cagattacat tgcacaaaga cagaactgag cttctctagc tagagtcgat
     1921 cgacaagctc gagtttctcc ataataatgt gtgagtagtt cccagataag ggaattaggg
     1981 ttcctatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     2041 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa
     2101 tccagatcgc tactattgtc ttctgcacga agtggtttaa actatcagtg tttgacagga
     2161 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     2221 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     2281 cagatcaggc gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
     2341 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
     2401 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
     2461 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
     2521 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
     2581 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
     2641 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
     2701 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
     2761 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     2821 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     2881 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     2941 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     3001 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     3061 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     3121 cggcctgctt ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     3181 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     3241 gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     3301 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact
     3361 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     3421 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     3481 ttcaccctcc ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat
     3541 ttgacccgcc tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc
     3601 cgtagaccgt ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata
     3661 ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga
     3721 tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc
     3781 tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4681 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     4741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5461 aagcatttat caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5521 taaacaaata ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg
     5581 ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag
     5641 ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc
     5701 ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg
     5761 gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc
     5821 ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga
     5881 tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat
     5941 ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat
     6001 ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa
     6061 cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag
     6121 ggccaagttt tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt
     6181 cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc gccagccc
//
