LOCUS       sequence-349540-        6628 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MoClo Level 1, position 2, transcriptional unit for transient 
            expression of 7-deoxyloganic acid hydroxylase (Cr7-DLH) 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 6628)
  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 6628)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pEPQD1CB0804
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    117..172
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      1741..1993
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    2039..2063
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2219..3367)
                     /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(3703..4291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4462..5253,5254..5322))
                     /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     5085..5104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5323..5427)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5514..6145
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6237..6261
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6378..6411
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6437..6470
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6500..6533
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6559..6592
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        join(6609..6628,1..27)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
ORIGIN
        1 aaggaagttc atttcatttg gagaggactc cggtattttt acaacaatta ccacaacaaa
       61 acaaacaaca aacaacatta caatttacta ttctagtcga atactgtatt tttacaacaa
      121 ttaccaacaa caacaaacaa caaacaacat tacaattact atttacaatt acaatggaat
      181 tgaacttcaa atcaattatt ttcttggttt ttgttagtct aaccctttac tgggtctata
      241 gaatcttgga ttgggtatgg tttaagccca agaagctgga aaaatgctta agagaacaag
      301 ggtttaaggg aaatccttac agattgttct tgggagatca gtatgatagt ggaaaattga
      361 tcagacaagc cttgactaaa cctattggtg ttgaagaaga tgttaagaaa cgaattgtac
      421 ctcatatact caaaactgtt gggactcatg gcaagaagtc ttttatgtgg gttggaagaa
      481 taccaagggt gaatatcaca gatccagagc tgattaaaga ggtcttaaca aaatactata
      541 agttccagaa gaatcatcat gatcttgacc caattaccaa attgcttttg actggaattg
      601 gaagcttaga aggtgaccct tgggctaaac gccgaaaaat catcaatgct gcttttcact
      661 ttgaaaagtt aaagcttatg cttcctgcat tctatttgag ttgccgtgat atggtgacaa
      721 aatgggataa taaggtccct gaaggaggat cagcagaggt ggatgtgtgg catgatattg
      781 aaaccttaac tggagatgta atttcaagaa cattatttgg aagtaacttc gaggaaggga
      841 gaagaatatt tgaactcatg aaagaactta ctgctcttac tattgatgtc attcgctcgg
      901 tttatatccc tggacagagg tttcttccaa ctaagaggaa caataggatg agagcaattg
      961 acaaagaagt tagagtaaga ataacagaaa ttatcaataa gaaaatgaag gtaatgaaat
     1021 caggagaagc agcaagtgct gctgatgatt ttttgggaat attgttggaa tgcaatctga
     1081 atgaaataaa ggagcaagga aataacaaaa gtgctggaat gactattgga gagattattg
     1141 gtgagtgcaa actcttctat tttgctggcc aggacactac ctcgacttta ctcgtgtgga
     1201 caatggtcct cttgtctaga ttccccgaat ggcaaactcg tgccagagaa gaagtttttc
     1261 aagtctttgg taataaaacc ccggactatg atggtatcag ccatcttaaa gttataacga
     1321 tgatcttgta cgaggttttg aggttatata ctccagtggc tgaactgaca aaagtggctc
     1381 atgaagctac acaactggga aaatacttca ttccagcggg cgtgcagcta atgatgccac
     1441 aaatcctact tcatcatgac cctgaaatat ggggtgaaga tgtaatggag ttcaaaccag
     1501 aaagatttgc agagggagtt ctcaaggcaa ctaagagcca aggctctttt tttccattta
     1561 gcttgggacc aagaatgtgc attggccaaa actttgctct attggaagcc aaaatggcaa
     1621 tgtctctaat cttgcgtaga ttttctttcg agctttctcc ctcctatgtt catgctcctt
     1681 tcactctcat cacaatgcaa ccccaatatg gtgctcattt gattctgcac aaactctaag
     1741 cttggactcc catgttggca aaggcaacca aacaaacaat gaatgatccg ctcctgcata
     1801 tggggcggtt tgagtatttc aactgccatt tgggctgaat tgtagacatg ctcctgtcag
     1861 aaattccgtg atcttactca atattcagta atctcggcca atatcctaaa tgtgcgtggc
     1921 tttatctgtc tttgtattgt ttcatcaatt catgtaacgt ttgcttttct tatgaatttt
     1981 caaataaatt atccgctact attgtcttct gcacgaagtg gtttaaacta tcagtgtttg
     2041 acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat
     2101 ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg
     2161 gttccccaga tcaggcgctg gctgctgaac ccccagccgg aactgacccc acaaggccct
     2221 agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg ccgctgcctc
     2281 gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg gtggaatccg
     2341 atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg tgcgagctga
     2401 aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc catatgaatt
     2461 tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg acctggcaac
     2521 gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac accgattcca
     2581 ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt
     2641 cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg caaagctcgt
     2701 agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc aacacctgct
     2761 gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt
     2821 ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc
     2881 gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc
     2941 acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc gtgtgtttca
     3001 gcaacgcggc ctgcttggcc tcgctgacct gttttgccag gtcctcgccg gcggtttttc
     3061 gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg
     3121 cctcctgttc aagacgacgc gaacgctcca cggcggccga tggcgcgggc agggcagggg
     3181 gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggacc atcgagccga
     3241 cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat
     3301 cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc
     3361 gattcattca ccctccttgc gggattgccc cgactcacgc cggggcaatg tgcccttatt
     3421 cctgatttga cccgcctggt gccttggtgt ccagataatc caccttatcg gcaatgaagt
     3481 cggtcccgta gaccgtctgg ccgtccttct cgtacttggt attccgaatc ttgccctgca
     3541 cgaataccag cgaccccttg cccaaatact tgccgtgggc ctcggcctga gagccaaaac
     3601 acttgatgcg gaagaagtcg gtgcgctcct gcttgtcgcc ggcatcgttg cgccacatct
     3661 aggatctgcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc
     4621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5341 ttattgaagc atttatcaag gttattgtct catgagcgga tacatatttg aatgtattta
     5401 gaaaaataaa caaatagggg ttccgcgcac gaattggcca gcgctgccat ttttggggtg
     5461 aggccgttcg cggccgaggg gcgcagcccc tggggggatg ggaggcccgc gttagcgggc
     5521 cgggagggtt cgagaagggg gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc
     5581 gcagccctgg ttaaaaacaa ggtttataaa tattggttta aaagcaggtt aaaagacagg
     5641 ttagcggtgg ccgaaaaacg ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga
     5701 cagcccctca aatgtcaata ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt
     5761 caaggatcgc gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc
     5821 acttatcccc aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc aggcgttttc
     5881 gccgatttgc gaggctggcc agctccacgt cgccggccga aatcgagcct gcccctcatc
     5941 tgtcaacgcc gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc
     6001 agtgagggcc aagttttccg cgaggtatcc acaacgccgg cggccgcggt gtctcgcaca
     6061 cggcttcgac ggcgtttctg gcgcgtttgc agggccatag acggccgcca gcccagcggc
     6121 gagggcaacc agcccggtga gcgtcgcaaa ggagatcctg atctgactga tgggctgcct
     6181 gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc
     6241 aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg
     6301 tttttaatgt actggggtgg atgcagtggg ccccactctg tgaagacaag caagaattca
     6361 agcttggagc ggtgcgggtg ccagggcgtg cccttgggct ccccgggcgc gtactccact
     6421 ccccgagccg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccactc
     6481 cccccatggt gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca
     6541 ctccccgagc cggtgcgggt gccagggcgt gcccttgggc tccccgggcg cgtactccac
     6601 tccccttcgc aagacccttc ctctatat
//
