LOCUS       sequence-404854-        8346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of the CrGLK coding sequence from C. roseus in a 
            plant binary vector for Agrobacterium-mediated transformation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8346)
  AUTHORS   Cole-Osborn LF, McCallan SA, Prifti O, Abu R, Sjoelund V, 
            Lee-Parsons CWT
  TITLE     The role of the Golden2-like (GLK) transcription factor in 
            regulating terpenoid indole alkaloid biosynthesis in Catharanthus 
            roseus.
  JOURNAL   Plant Cell Rep. 2024 May 14;43(6):141. doi: 
            10.1007/s00299-024-03208-9.
  PUBMED    38743349
REFERENCE   2  (bases 1 to 8346)
  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: "Plant Cell 
            Rep."; date: "2024-05-14"; volume: "43(6):141. doi"; pages: " 
            10.1007/s00299-024-03208-9"
COMMENT     Sequence Label: pLC047 - pL2_p2x35s-CrGLK-tMAS
FEATURES             Location/Qualifiers
     source          1..8346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     244..264
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      1764..2016
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    2080..2104
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2384..3013
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3442..4515
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      4581..4775
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4837..4941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4942..5736
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7061..7649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7550..7569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7785..7809
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(8302..8346,1..301)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta
       61 caaatgccat cattgcgata aaggaaaggc tatcattcaa gatctctctg ccgacagtgg
      121 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagagg ttccaaccac
      181 gtctacaaag caagtggatt gatgtgacat ctccactgac gtaagggatg acgcacaatc
      241 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      301 acgctcgagt ataagagctc atttttacaa caattaccaa caacaacaaa caacaaacaa
      361 cattacaatt acatttacaa ttatcgatac aatgcttgct gtgtcaccat tgaggagcaa
      421 taataatgag agaaaaaagg agatatgtga aattagtagt agtactagtt ttccagttgt
      481 tgatgatttt cctgattttg ggggagcaaa tttgcttgat agcattgatt ttgatgaact
      541 ttttgttggg attaacggtg aggatgatgt tttaccagat ttggaaatgg atcctcatga
      601 tattcttgct gaattctcac ttactgaatc attatcacca tctgtttctc ttgatcatca
      661 acatcatgaa cattctcaaa ttacctcctc aatttcggcg gctgattatg ataaactagt
      721 agaattagag gataaggcta attcatcatc cgcttcagac gacgtcggtt caggctcatc
      781 tactttaact caaggagaag atgttgttag caaatcaacg gttttaaatc catcttctaa
      841 agaaggtgat aaaggaagga agtcatcttc tactcaatca aagaattctt ctgggaagaa
      901 gaaagtgaag gtggattgga caccagagct gcacagaaga tttgtacaag cagtggagca
      961 gctaggtgtg gataaggcag tcccttcaag aattcttgaa cttatgggaa ttgattgtct
     1021 tactcgccac aacattgcta gccatcttca aaaatacaga tcccacagga aacatttgct
     1081 agcaagagaa gctgaggcag caaattggag ccagaggcgg caattatacg gaggcactgc
     1141 agcagcagca ggaggaggga agagagatgt aaatccatgg cttgctccaa ccatgggatt
     1201 tcctcctccc atggcaccgg cggccgtccc tcattttaga cccttacatg tatggggtca
     1261 cccatctgtc gaccaatctt taatgcacgt ttggcctaaa catttagctc ctccgccccg
     1321 accaccaaca tggccaccaa ctccaacacc aacaccaaca ccaccaccac caccgccaga
     1381 tcctaattcc ttttggcatt cacctcacca acgtgtacca aaccctctta ccccaggaac
     1441 gccttgcttt ccgccacctt taacaccagc gagatttcca acaccgccgg tccccggaat
     1501 accaccacct ccaatgtaca aagtagaccc ggtcgcaggc atgggtgtgc cggcacctgc
     1561 taccggacaa gcagtacccc atgcacctcc tcttgacttt cacccctcga cggagagcgt
     1621 agatgcagct atcggagatg ttttatcgaa accctggctg ccacttcctc ttgggctgaa
     1681 accgccgtcg attgacagtg tactggtgga gctacagcgg caaggaatac cgaaaattcc
     1741 accagtttgt acgggttcgt aagcttggac tcccatgttg gcaaaggcaa ccaaacaaac
     1801 aatgaatgat ccgctcctgc atatggggcg gtttgagtat ttcaactgcc atttgggctg
     1861 aattgtagac atgctcctgt cagaaattcc gtgatcttac tcaatattca gtaatctcgg
     1921 ccaatatcct aaatgtgcgt ggctttatct gtctttgtat tgtttcatca attcatgtaa
     1981 cgtttgcttt tcttatgaat tttcaaataa attatccgct gcaagaggat gcacatgtga
     2041 ccgagggaca cgaagtgatc cgtttaaact atcagtgttt gacaggatat attggcgggt
     2101 aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg
     2161 tttatccgtt cgtccatttg tatgtgccag ccgtgcggct gcatgaaatc ctggccggtt
     2221 tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc
     2281 gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt
     2341 atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt
     2401 acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct
     2461 gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga
     2521 ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat
     2581 tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca
     2641 ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca
     2701 gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat
     2761 tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac
     2821 gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc
     2881 ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga
     2941 atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc
     3001 aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt
     3061 gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg
     3121 ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg
     3181 tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta
     3241 ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg
     3301 catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac
     3361 gggcggttgg ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg gcactggaac
     3421 ccccaagccc gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc
     3481 gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa
     3541 cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc
     3601 aaagaatccc ggcaaccgcc ggcagccggt gcgtcgtcga ttaggaagcc gcccaagggc
     3661 gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg cgatagtcgc
     3721 agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg
     3781 atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc cggcatggcc
     3841 agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga atccatgaac
     3901 cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca cgttgcggac
     3961 gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct ggtagaaacc
     4021 tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc caagaacggc
     4081 cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat cgtaaagagc
     4141 gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta ccgcgagatc
     4201 acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt gatcgatccc
     4261 ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc agaagccaga
     4321 tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt
     4381 ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt gaaggaggag
     4441 gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga gggcgaagca
     4501 tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa
     4561 ggtcgaaaaa gcttctttcc tgtggatagc acgtacattg ggaacccaaa gccgtacatt
     4621 gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg gtcacacatg
     4681 taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc tttaaaactt
     4741 attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg cacagccgaa
     4801 cagctgcaaa aagcgcccac atcaaggctc cgagtgcgcg gaacccctat ttgtttattt
     4861 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4921 taatattgaa aaaggaagag tatggctaaa atgagaatat caccggaatt gaaaaaactg
     4981 atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa ggtatataag
     5041 ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta taaagggacc
     5101 acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa gctgcctgtt
     5161 ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat gagtgaggcc
     5221 gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa gattatcgag
     5281 ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg tccctatacg
     5341 aatagcttag acagccgctt agccgaattg gattacttac tgaataacga tctggccgat
     5401 gtggattgcg aaaactggga agaggacact ccatttaaag atccgcgcga gctgtatgat
     5461 tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga cctgggagac
     5521 agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg gagaagcggc
     5581 agggcggaca agtggtatga cattgccttc tgcgtccggt cgctcaggga ggatatcggg
     5641 gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga ttgggagaaa
     5701 ataaaatatt atattttact ggatgaattg ttttagctgt cagaccaagt ttactcatat
     5761 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5821 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttctagct gtaacctcga
     5881 agcgtttcac ttgtaacaac gattgagaat ttttgtcata aaattgaaat acttggttcg
     5941 catttttgtc atccgcggtc agccgcaatt ctgacgaact gcccatttag ctggagatga
     6001 ttgtacatcc ttcacgtgaa aatttctcaa gcgctgtgaa caagggttca gattttagat
     6061 tgaaaggtga gccgttgaaa cacgttcttc ttgtcgatga cgacgtcgct atgcggcatc
     6121 ttattattga ataccttacg atccacgcct tcaaagtgac cgcggtagcc gacagcaccc
     6181 agttcacaag agtactctct tccgcgacgg tcgatgtcgt ggttgttgat ctagatttag
     6241 gtcgtgaaga tgggctcgag atcgttcgta atctggcggc aaagtctgat attccaatca
     6301 taattatcag tggcgaccgc cttgaggaga cggataaagt tgttgcactc gagctaggag
     6361 caagtgattt tatcgctaag ccgttcagta tcagagagtt tctagcacgc attcgggttg
     6421 ccttgcgcgt gcgccccaac gttgtccgct ccaaagaccg acggtctttt tgttttactg
     6481 actggacact taatctcagg caacgtcgct tgatgtccga agctggcggt gaggtgaaac
     6541 ttacggcagg tgagttcaat cttctcctcg cgtttttaga gaaaccccgc gacgttctat
     6601 cgcgcgagca acttctcatt gccagtcgag tacgcgacga ggaggtttat gacaggagta
     6661 tagatgttct cattttgagg ctgcgccgca aacttgaggc ggatccgtca agccctcaac
     6721 tgataaaaac agcaagaggt gccggttatt tctttgacgc ggacgtgcag gtttcgcacg
     6781 gggggacgat ggcagcctga gccaattgca tttgcctctt aattatctgg ctcaaagggt
     6841 gactgaggag taagcgatgt gcccatcaca ctgaccacca agacgggggt tcgttttgaa
     6901 gatcaccctt cagacccaag gccagtgtgc gttgatcgcg gccacactgc aaattcacat
     6961 actgtcgata aaggggcgtt ggacgagaac ccattcgtca ggatactggc tcgtgatctc
     7021 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7081 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7141 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7201 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7261 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7321 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7381 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7441 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7501 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7561 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7621 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7681 tgctcggatc tgttggaccg gacagtagtc atggttgatg ggctgcctgt atcgagtggt
     7741 gattttgtgc cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt
     7801 ggtgtaaaca aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac
     7861 tggggttgaa cactctgtgc cgaattcgga tccggaggtc aacatggtgg agcacgacac
     7921 tctggtctac tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg ctattgagac
     7981 ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca
     8041 cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa
     8101 aggaaaggct atcattcaag atctctctgc cgacagtggt cccaaagatg gacccccacc
     8161 cacgaggagc atcgtggaaa aagaagaggt tccaaccacg tctacaaagc aagtggattg
     8221 atgtgataac atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt
     8281 ctcagaagat caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct
     8341 cggatt
//
