LOCUS       sequence-349015-        6051 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6051)
  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 6051)
  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: pEPQD1CB0108
FEATURES             Location/Qualifiers
     source          1..6051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    155..179
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        365..710
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     653..673
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1789..1963
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2008..2032
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2188..3336)
                     /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(3672..4260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3752..3771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4431..5222,5223..5291))
                     /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     5054..5073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5292..5396)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5483..6051,1..63)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag cccggtgagc
       61 gtcgcaaagg agatcctgat ctgactgatg ggctgcctgt atcgagtggt gattttgtgc
      121 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
      181 aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac tggggtggat
      241 gcagtgggcc ccactctgtg aagacaagca agaattcaag cttggaggtc aacatggtgg
      301 agcacgacac tctggtctac tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg
      361 ctattgagac ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag
      421 ctatctgtca cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc
      481 attgcgataa aggaaaggct atcattcaag atctctctgc cgacagtggt cccaaagatg
      541 gacccccacc cacgaggagc atcgtggaaa aagaagaggt tccaaccacg tctacaaagc
      601 aagtggattg atgtgacatc tccactgacg taagggatga cgcacaatcc cactatcctt
      661 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca cgctcgagta
      721 taagagctca tttttacaac aattaccaac aacaacaaac aacaaacaac attacaatta
      781 catttacaat tatcgataca atgtgctcaa acacaaatgc ccagttggcc aatttgtctg
      841 aacaagtaaa ggggaaggtc actgaatttg atttcaagga gtacatgcgt tccaaggcaa
      901 tgtcagtgaa tgaggcactg gacagggctg tcccacttcg ttatcctgaa aaaatacatg
      961 aagctatgag gtattctctt ctagcaggag ggaagcgagt tcgtcctatt ctgtgcattg
     1021 cagcatgtga gcttgtggga gggagcgagg agctcgccat gccaactgcc tgtgcaatgg
     1081 aaattatcca cacaatgtct ttgattcatg atgacttgcc tcccatggac aatgatgatt
     1141 tacgccgagg taagcctaca aaccacaagg tgttcggtga aggcactgct gttcttgcgg
     1201 gggatgcact tctttcattt gcatttgagc acattgcagt gtccacaagc aagactgtgg
     1261 agtctgatag ggttttgagg gtggtatctg aattgggtag agcaataggc tcggaagggg
     1321 ttgcaggtgg gcaggttgct gatattacca gtcaggggaa tccttctgtt ggccttgaga
     1381 ctctggaatg gattcacatt cacaagactg cagtgctctt ggagtgctcc gttgctagtg
     1441 gggcgatcat cggtggtgct tctgatgatg agattgagag agttcgaaag tatgcccgtt
     1501 gcgtggggct tctgtttcag gttgtggatg acattctcga tgtcacgaaa tcatcagaag
     1561 aattgggtaa gactgcagca aaggatttgc ttagtgataa ggccacttat cccaagctca
     1621 tgggtttgga gaaagcaaag gagtttgctg atgaattgtt gggcaaagct aaggaggagt
     1681 tatctttctt caacccaact aaggctgcac ctctgttggg tcttgcagat tacattgcac
     1741 aaagacagaa ctgagcttct ctagctagag tcgatcgaca agctcgagtt tctccataat
     1801 aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc gctcatgtgt
     1861 tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa
     1921 tttctaattc ctaaaaccaa aatccagtac taaaatccag atcgctacta ttgtcttctg
     1981 cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga
     2041 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg
     2101 tccatttgta tgtgcatgcc aaccacaggg ttccccagat caggcgctgg ctgctgaacc
     2161 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
     2221 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
     2281 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
     2341 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
     2401 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
     2461 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
     2521 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
     2581 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
     2641 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
     2701 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
     2761 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
     2821 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
     2881 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
     2941 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct cgctgacctg
     3001 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
     3061 gatggtcatc gacttcgcca aacctgccgc ctcctgttca agacgacgcg aacgctccac
     3121 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
     3181 ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
     3241 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
     3301 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
     3361 gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg ccttggtgtc
     3421 cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc cgtccttctc
     3481 gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc ccaaatactt
     3541 gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg tgcgctcctg
     3601 cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa aaaggccgcg
     3661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4561 atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaagg ttattgtctc
     5341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcacg
     5401 aattggccag cgctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct
     5461 ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc
     5521 ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat
     5581 attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc
     5641 ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct
     5701 catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc
     5761 gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt
     5821 gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc
     5881 gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct
     5941 caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca
     6001 caacgccggc ggccgcggtg tctcgcacac ggcttcgacg gcgtttctgg c
//
