LOCUS       sequence-349533-        6672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 3, transcriptional unit for transient 
            expression of geranyl pyrophosphate synthase (PaGPPS1) from Picea 
            abies promoter regions contains 4x attB sites for recruitment fo 
            gal4AD-phiC31.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  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 6672)
  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: pEPQD1CB0786
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    341..365
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    482..515
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    541..574
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    604..637
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    663..696
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        713..759
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      2059..2766
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2815..2839
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2995..4143)
                     /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(4479..5067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4559..4578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5238..6029,6030..6098))
                     /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     5861..5880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6099..6203)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6290..6672,1..249)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg
       61 ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt
      121 tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt
      181 tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg
      241 gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt
      301 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg
      361 taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg
      421 gtggatgcag tgggccccac tctgtgaaga caaactagaa ttcgagctcg gagcggtgcg
      481 ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cactccccga gccggtgcgg
      541 gtgccagggc gtgcccttgg gctccccggg cgcgtactcc actcccccca tggtgcggtg
      601 cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccactcccc gagccggtgc
      661 gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccactcccct tcgcaagacc
      721 cttcctctat ataaggaagt tcatttcatt tggagaggac tccggtattt ttacaacaat
      781 taccacaaca aaacaaacaa caaacaacat tacaatttac tattctagtc gaatactgta
      841 taagagctca tttttacaac aattaccaac aacaacaaac aacaaacaac attacaatta
      901 catttacaat tatcgataac atggcttctt ctatgctttc ttctgctgct gttgttgcta
      961 ctcgtgctag tgctgctcaa gctagtatgg ttgctccttt tactggactt aagtctgctg
     1021 cttcttttcc tgttactaga aagcaaaaca accttgatat tacttctatt gctagtaacg
     1081 gaggaagagt tcgagcaatg tgctcaaaca caaatgccca gttggccaat ttgtctgaac
     1141 aagtaaaggg gaaggtcact gaatttgatt tcaaggagta catgcgttcc aaggcaatgt
     1201 cagtgaatga ggcactggac agggctgtcc cacttcgtta tcctgaaaaa atacatgaag
     1261 ctatgaggta ttctcttcta gcaggaggga agcgagttcg tcctattctg tgcattgcag
     1321 catgtgagct tgtgggaggg agcgaggagc tcgccatgcc aactgcctgt gcaatggaaa
     1381 ttatccacac aatgtctttg attcatgatg acttgcctcc catggacaat gatgatttac
     1441 gccgaggtaa gcctacaaac cacaaggtgt tcggtgaagg cactgctgtt cttgcggggg
     1501 atgcacttct ttcatttgca tttgagcaca ttgcagtgtc cacaagcaag actgtggagt
     1561 ctgatagggt tttgagggtg gtatctgaat tgggtagagc aataggctcg gaaggggttg
     1621 caggtgggca ggttgctgat attaccagtc aggggaatcc ttctgttggc cttgagactc
     1681 tggaatggat tcacattcac aagactgcag tgctcttgga gtgctccgtt gctagtgggg
     1741 cgatcatcgg tggtgcttct gatgatgaga ttgagagagt tcgaaagtat gcccgttgcg
     1801 tggggcttct gtttcaggtt gtggatgaca ttctcgatgt cacgaaatca tcagaagaat
     1861 tgggtaagac tgcagcaaag gatttgctta gtgataaggc cacttatccc aagctcatgg
     1921 gtttggagaa agcaaaggag tttgctgatg aattgttggg caaagctaag gaggagttat
     1981 ctttcttcaa cccaactaag gctgcacctc tgttgggtct tgcagattac attgcacaaa
     2041 gacagaactg agcttgtcct gctttaatga gatatgcgag aagcctatga tcgcatgata
     2101 tttgctttca attctgttgt gcacgttgta aaaaacctga gcatgtgtag ctcagatcct
     2161 taccgccggt ttcggttcat tctaatgaat atatcacccg ttactatcgt atttttatga
     2221 ataatattct ccgttcaatt tactgattgt accctactac ttatatgtac aatattaaaa
     2281 tgaaaacaat atattgtgct gaataggttt atagcgacat ctatgataga gcgccacaat
     2341 aacaaacaat tgcgttttat tattacaaat ccaattttaa aaaaagcggc agaaccggtc
     2401 aaacctaaaa gactgattac ataaatctta ttcaaatttc aaaagtgccc caggggctag
     2461 tatctacgac acaccgagcg gcgaactaat aacgctcact gaagggaact ccggttcccc
     2521 gccggcgcgc atgggtgaga ttccttgaag ttgagtattg gccgtccgct ctaccgaaag
     2581 ttacgggcac cattcaaccc ggtccagcac ggcggccggg taaccgactt gctgccccga
     2641 gaattatgca gcattttttt ggtgtatgtg ggccccaaat gaagtgcagg tcaaaccttg
     2701 acagtgacga caaatcgttg ggcgggtcca gggcgaattt tgcgacaaca tgtcgaggct
     2761 cagcaggacc gctttacttg tcttctgcac gaagtggttt aaactatcag tgtttgacag
     2821 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     2881 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     2941 cccagatcag gcgctggctg ctgaaccccc agccggaact gaccccacaa ggccctagcg
     3001 tttgcaatgc accaggtcat cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa
     3061 ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc
     3121 gcacatgagg cggaaggttt ccagcttgag cgggtacggc tcccggtgcg agctgaaata
     3181 gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac ttctcccata tgaatttcgt
     3241 gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct ggcaacggga
     3301 cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg attccaggtg
     3361 cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc
     3421 ggccttcgtg taataccggc cattgatcga ccagcccagg tcctggcaaa gctcgtagaa
     3481 cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg tactccaaca cctgctgcca
     3541 caccagttcg tcatcgtcgg cccgcagctc gacgccggtg taggtgatct tcacgtcctt
     3601 gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt
     3661 ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg
     3721 cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa
     3781 cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt
     3841 cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc
     3901 ctgttcaaga cgacgcgaac gctccacggc ggccgatggc gcgggcaggg cagggggagc
     3961 cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc tggaccatcg agccgacgga
     4021 ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt cggcatcctc
     4081 ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt
     4141 cattcaccct ccttgcggga ttgccccgac tcacgccggg gcaatgtgcc cttattcctg
     4201 atttgacccg cctggtgcct tggtgtccag ataatccacc ttatcggcaa tgaagtcggt
     4261 cccgtagacc gtctggccgt ccttctcgta cttggtattc cgaatcttgc cctgcacgaa
     4321 taccagcgac cccttgccca aatacttgcc gtgggcctcg gcctgagagc caaaacactt
     4381 gatgcggaag aagtcggtgc gctcctgctt gtcgccggca tcgttgcgcc acatctagga
     4441 tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc accggctcca
     5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6121 tgaagcattt atcaaggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6181 aataaacaaa taggggttcc gcgcacgaat tggccagcgc tgccattttt ggggtgaggc
     6241 cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta gcgggccggg
     6301 agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca cagggcgcag
     6361 ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa gacaggttag
     6421 cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc tgtggacagc
     6481 ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc aagtgtcaag
     6541 gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg cagggcactt
     6601 atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc gttttcgccg
     6661 atttgcgagg ct
//
