LOCUS       sequence-349545-        7008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 6, transcriptional unit for transient 
            expression of geraniol 8-oxidase (CrG8H) from Catharanthus roseus 
            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 7008)
  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 7008)
  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: pEPQD1CB0865
FEATURES             Location/Qualifiers
     source          1..7008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..41
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    71..104
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    130..163
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        180..226
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    316..371
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      1862..2569
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2618..2642
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2798..3946)
                     /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(4282..4870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4362..4381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5041..5832,5833..5901))
                     /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     5664..5683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5902..6006)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6093..6724
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6816..6840
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6957..6990
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
ORIGIN
        1 ggtgcgggtg ccagggcgtg cccttgggct ccccgggcgc gtactccact ccccccatgg
       61 tgcggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc actccccgag
      121 ccggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca ctccccttcg
      181 caagaccctt cctctatata aggaagttca tttcatttgg agaggactcc ggtattttta
      241 caacaattac cacaacaaaa caaacaacaa acaacattac aatttactat tctagtcgaa
      301 tactgtattt ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacta
      361 tttacaatta caatggatta ccttaccata atattaactt tactatttgc cttgactctc
      421 tatgaagcct tcagctacct atccagaaga accaaaaacc ttcctccagg accatcgcca
      481 ttgccgttca tcggaagcct ccatttatta ggcgaccaac cacacaaatc cttagcaaaa
      541 ctttccaaaa aacacggtcc aattatgagt ctcaaattag gccagatcac tacaatcgtc
      601 atatcttcat caacaatggc gaaagaagtt cttcaaaaac aggatttagc attttcaagc
      661 agatcagttc caaacgcact ccacgctcac aatcaattca aattctccgt tgtatggctt
      721 ccggtagcct cacgatggag aagccttcga aaagttttga attctaatat attttccggc
      781 aatcggctcg acgctaatca acatttgaga actagaaaag tacaggaact aattgcgtat
      841 tgccggaaaa atagccagag cggagaagcg gttgacgtcg gccgagctgc ttttagaact
      901 tcgttgaatt tgttgtcgaa tttgattttt tcaaaggatt tgacggatcc ttattcggat
      961 tctgccaagg aattcaagga tttggtttgg aatataatgg ttgaggcggg gaaacctaat
     1021 ttggtcgatt tttttcccct gcttgaaaaa gttgatcctc aaggtatacg acatcgtatg
     1081 acgattcact ttggggaagt tcttaagctt tttggtggac ttgttaatga aagattggag
     1141 caaagaagat caaaagggga aaaaaatgat gtgttggatg tacttctaac taccagccaa
     1201 gaaagccctg aggaaatcga tagaactcac attgagcgaa tgtgcttgga cctgtttgta
     1261 gcagggacgg acacaacatc aagcacatta gaatgggcaa tgtcagaaat gcttaaaaac
     1321 ccagacaaaa tgaagaaaac ccaagatgaa cttgcacaag taatcggcag aggaaaaaca
     1381 atagaagaat ccgatattaa ccgcttacct tacttaagat gcgttatgaa agaaacctta
     1441 aggatacatc caccagttcc cttcttaatt cctcgcaaag tggaacaaag tgttgaggtt
     1501 tgtggataca atgtccctaa aggatcacaa gttcttgtga atgcttgggc aattggacgt
     1561 gatgaaactg tttgggatga tgctttggca ttcaaacccg agagatttat ggaatctgaa
     1621 ttggatatcc gtggaagaga tttcgagctg attccgttcg gtgctggccg aagaatttgc
     1681 ccagggttgc cattggcact aaggactgtg cctttgatgc ttggttcttt gttgaactct
     1741 tttaattgga agcttgaagg tgggatggct ccaaaagatt tggatatgga ggagaagttt
     1801 ggtattacac tgcagaaggc tcatcctttg cgtgctgtac caagcaccct ttaagcttgt
     1861 cctgctttaa tgagatatgc gagaagccta tgatcgcatg atatttgctt tcaattctgt
     1921 tgtgcacgtt gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc ggtttcggtt
     1981 cattctaatg aatatatcac ccgttactat cgtattttta tgaataatat tctccgttca
     2041 atttactgat tgtaccctac tacttatatg tacaatatta aaatgaaaac aatatattgt
     2101 gctgaatagg tttatagcga catctatgat agagcgccac aataacaaac aattgcgttt
     2161 tattattaca aatccaattt taaaaaaagc ggcagaaccg gtcaaaccta aaagactgat
     2221 tacataaatc ttattcaaat ttcaaaagtg ccccaggggc tagtatctac gacacaccga
     2281 gcggcgaact aataacgctc actgaaggga actccggttc cccgccggcg cgcatgggtg
     2341 agattccttg aagttgagta ttggccgtcc gctctaccga aagttacggg caccattcaa
     2401 cccggtccag cacggcggcc gggtaaccga cttgctgccc cgagaattat gcagcatttt
     2461 tttggtgtat gtgggcccca aatgaagtgc aggtcaaacc ttgacagtga cgacaaatcg
     2521 ttgggcgggt ccagggcgaa ttttgcgaca acatgtcgag gctcagcagg accgctgagc
     2581 ttgtcttctg cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa
     2641 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     2701 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat caggcgctgg
     2761 ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt
     2821 catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc
     2881 cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg
     2941 tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg
     3001 ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa
     3061 acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt
     3121 ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg
     3181 tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc
     3241 ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct
     3301 cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt
     3361 cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga
     3421 ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc
     3481 gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg
     3541 aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct
     3601 cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc
     3661 ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca agacgacgcg
     3721 aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc
     3781 gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg
     3841 gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt
     3901 cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg
     3961 ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg
     4021 ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc
     4081 cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc
     4141 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
     4201 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
     4261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4681 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5161 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     5221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaagg
     5941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6001 tccgcgcacg aattggccag cgctgccatt tttggggtga ggccgttcgc ggccgagggg
     6061 cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
     6121 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
     6181 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     6241 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     6301 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     6361 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     6421 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     6481 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     6541 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
     6601 gaggtatcca caacgccggc ggccgcggtg tctcgcacac ggcttcgacg gcgtttctgg
     6661 cgcgtttgca gggccataga cggccgccag cccagcggcg agggcaacca gcccggtgag
     6721 cgtcgcaaag gagatcctga tctgactgat gggctgcctg tatcgagtgg tgattttgtg
     6781 ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac
     6841 aaattgacgc ttagacaact taataacaca ttgcggacgt ttttaatgta ctggggtgga
     6901 tgcagtgggc cccactctgt gaagacaatg tggaattcct cgagggagcg gtgcgggtgc
     6961 cagggcgtgc ccttgggctc cccgggcgcg tactccactc cccgagcc
//
