LOCUS       sequence-345738-        5473 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway (Invitrogen) entry clone (pDONR207) containing the MUM2 
            (At5g63800) signal peptide (84bp) fused in-frame to the Citrine and 
            the coding sequence of the HG degrading enzyme ADPG2 (At2g41850.1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5473)
  AUTHORS   McGee R, Dean GH, Wu D, Zhang Y, Mansfield SD, Haughn GW
  TITLE     Pectin Modification in Seed Coat Mucilage by In Vivo Expression of 
            Rhamnogalacturonan-I- and Homogalacturonan-Degrading Enzymes.
  JOURNAL   Plant Cell Physiol. 2021 Jun 1. pii: 6290394. doi: 
            10.1093/pcp/pcab077.
  PUBMED    34059917
REFERENCE   2  (bases 1 to 5473)
  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: "Plant Cell 
            Physiol. 2021 Jun 1. pii: 6290394. doi: 10.1093/pcp/pcab077."
COMMENT     Sequence Label: pDONR207-MUM2sp-Citrine-ADPG2
FEATURES             Location/Qualifiers
     source          1..5473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(73..100)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(192..278)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    331..430
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(542..544,545..1255)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(584..605)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(845..864)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1192..1213
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2534..2633)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2745..2764)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2823..2842)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3416..3949)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4138..4143,4144..4160,4161..4166))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagcc aggaagagtt
       61 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
      121 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
      181 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      241 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      301 cgcgttaacg ctagcatgga tctcgggccc caaataatga ttttattttg actgatagtg
      361 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
      421 aaagcaggct atggagatgg gtcgtctggt cttcgggttg tgtttgattc ttattgtggg
      481 tacgtttctt gaattctccg gcggtgctac ggcgctgcag cctggaggtg gaggtggagc
      541 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      601 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      661 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      721 cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca
      781 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      841 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      901 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      961 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1021 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1081 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1141 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1201 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggatcc
     1261 tgctggtgct gctgcgcctc tagcagcact agtgtcaaga atcagcccta atgtatatga
     1321 ccattcttat aaaaggttca aatccgatag cttaatcaag cgaagagagg acatcacggg
     1381 attgagaagt tttgtaagag cctctttgcg gactccaacc accgttagtg tttctgattt
     1441 tggagctaaa ggagatggaa aaaccgatga cacgcaggcg ttcgtgaatg cgtggaagaa
     1501 agcatgttct tcaaatggag ctgttaatct cctagttcct aaagggaata cttatctcct
     1561 taagtctatt caattaactg gtccatgcaa ttctattctc accgttcaga tattcggaac
     1621 gttatccgca tctcaaaaac gatcggatta caaagatatc agcaaatgga taatgtttga
     1681 tggcgttaac aatctatcag tcgatggagg cgacactggg gttgtggacg gaaacggcga
     1741 aacgtggtgg caaaactcat gcaaacggaa caaagctaag ccatgcacaa aagccccaac
     1801 ggctcttact ttttacaact cgaaaagtct gatagtgaag aatctgaagg tgagaaatgc
     1861 acagcagatc cagatttcga ttgaaaaatg ctccaacgtt caggtctcta atgtcgtggt
     1921 aactgcgcct gcggatagtc ctaacaccga tggtattcat atcactaaca cccaaaacat
     1981 tcgagtctcc gaatccatca ttggaacagg cgatgattgc atatctattg aaagtggatc
     2041 acaaaatgtt caaatcaatg atataacttg cggtcccggt cacggtatca gtattgggag
     2101 ccttggagat gacaattcaa aggcttttgt ctcaggcgtg actgtggatg gtgctaagct
     2161 ttccggtaca gacaatggag taagaatcaa aacttaccag ggagggtcag gaactgctag
     2221 caatattata tttcaaaaca ttcagatgga taatgttaag aatccgatca taatcgacca
     2281 agactactgc gacaagagca aatgcactac agagaaatcc gcggtccaag tgaagaacgt
     2341 ggtgtaccgg gacataagtg gcactagcgc atcggaaaac gcaataacgt ttaactgcag
     2401 caagaactat ccatgccaag gaattgtgct tgacagagtg aacattaaag gaggaaaagc
     2461 aacttgcacc aatgctaatg tggttgataa aggagctgtt ctgcctcagt gcaactccac
     2521 ttagggacgt cctacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     2581 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     2641 gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2701 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2761 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2821 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2881 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2941 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3001 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     3061 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3121 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3181 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3241 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3301 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3361 tatttttgac gaggggaaat taggccggga agccgatctc ggcttgaacg aattgttagg
     3421 tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat
     3481 agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa
     3541 gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt
     3601 gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg
     3661 gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga
     3721 tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt
     3781 aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga
     3841 cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg
     3901 ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc
     3961 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
     4021 gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
     4081 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt
     4141 ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg
     4201 cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac
     4261 gagcgcaagg tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac
     4321 ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg
     4381 cggcgcttgc cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa
     4441 ggcgagcatc gtttgttcgc ccaggactct agctatagtt ctagtggttg gctactaata
     4501 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     4561 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4621 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4681 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     4741 gcaagctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gtt
//
