LOCUS       sequence-356729-        8238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MUM2 signal peptide fused to Citrine, under control of MUM4_1.5kb 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8238)
  AUTHORS   Lee YC, Dean GH, Gilchrist E, Tsai AY, Haughn GW
  TITLE     Asymmetric distribution of extracellular matrix proteins in seed 
            coat epidermal cells of Arabidopsis is determined by polar 
            secretion.
  JOURNAL   Plant Direct. 2021 Nov 28;5(11):e360. doi: 10.1002/pld3.360. 
            eCollection 2021 Nov.
  PUBMED    34877448
REFERENCE   2  (bases 1 to 8238)
  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 
            Direct."; date: "2021-11-28"; volume: "5(11):e360. doi"; pages: " 
            10.1002/pld3.360. eCollection 2021 Nov"
COMMENT     Sequence Label: MUM2SP-Citrine
FEATURES             Location/Qualifiers
     source          1..8238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    625..647
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(671..923)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(943..1494)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1536..1715)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1947..1969
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1961..1978
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1962..1978
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1988..2007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1988..2006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3562..3564,3565..4275)
                     /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(3604..3625)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3865..3884)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4212..4233
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4591..4611)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4591..4609)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4630..4646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4630..4646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4643..4665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4654..4670
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4678..4684,4685..4702,4703..4708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4723..4744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4861..4878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4983..5007
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5098..5686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5178..5197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7195..8010)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7195..8010)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7918..7937
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8002..8021
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
       61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
      121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
      181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
      241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
      301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
      361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
      421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
      481 ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta
      541 taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga
      601 tttccgatcc ccggaattag atcttggcag gatatattgt ggtgtaacgt tatcagcttg
      661 catgccggtc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg
      721 cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa
      781 acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc
      841 aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt
      901 tattgccaaa tgtttgaacg atctgcttga ctctaggggt catcagattt cggtgacggg
      961 caggaccgga cggggcggca ccggcaggct gaagtccagc tgccagaaac ccacgtcatg
     1021 ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc
     1081 ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa
     1141 gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg
     1201 ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg cgttgcgtgc
     1261 cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca
     1321 gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc
     1381 ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg
     1441 catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcatggtaga
     1501 tcccctcgat cgagttgaga gtgaatatga gactctaatt ggataccgag gggaatttat
     1561 ggaacgtcag tggagcattt ttgacaagaa atatttgcta gctgatagtg accttaggcg
     1621 acttttgaac gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa
     1681 cccgcggctc agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct
     1741 tgtcccgcgt catcggcggg ggtcataacg tgactccctt aattctccgc tcatgatcga
     1801 taacattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     1861 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     1921 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     1981 gcgcgcgtaa tacgactcac tatagggcga attgggtacc aatcggtccc attagtcttt
     2041 attgttcttc aacaaaaaaa ggaatctctt tggttattgg aaaatatggt cttgattggt
     2101 taatatagac aacattatgc cgaagaatgt ggaagtaaca aaataaaaca ttggtcctta
     2161 gatttttctt ctgtaaacca ctctttctca gattatataa ttagtcaatt ttattcaatg
     2221 gttacaaaaa gaaaaaaaaa aacttgccaa tgaaaaaaag gaatgctctt gacctcaaga
     2281 ctcatcatgg ttaacaatga aattcgcctt aacttcgaaa ttttgacgcc gttgaaatgt
     2341 ttataattta cccgccaagt ttgtaggcct gagtttagcc cattaaaggt aatattaggc
     2401 gtccattaat cataacgaat atcactttct caatttaaca agtcatgaga aagtgtacac
     2461 atttgatcta tttaatattt attaaccaaa aaggaagcaa atcaattatt tcgttacgtg
     2521 acggtggcat taagcatctt gcattgaatg atccgttata tataatctca ggtttttttt
     2581 gggttgaaat gatgatatta aattttaggt tgacatgtac ttatctttgt aatcaactaa
     2641 ttaaatattt gaactgacat gtctacgtta tatcataaat aaaccaggtg ttttaattaa
     2701 ataccacgat taaccttcta aaataaggaa aatcatattt tattcgtcaa tcactataat
     2761 ttggaaaacg atgcaatata tttatttctt tctttataca catacttaat taattatcaa
     2821 aatttcattc tattttaatt gatctgaaat gtcctaattt agtagagagc taataaatag
     2881 actcacattt atgttgaaaa acaaaacaag aaagagaaaa agacatgagt ttaatgtcgg
     2941 acgctagact cacgagaaat ccgtaatatc taagacgatc aagggtaaat tagtcaattt
     3001 cctcctacgg ttctccgtaa ctgtctcaga atataagagg cacattgctg agagaagctc
     3061 cgtagaccgt agattccctc ttcttcttct tcctcttcct ctctactgtt ttttcttttc
     3121 atatttctct ctctctctct ctctatagtt aatgaagttt ctatctttat ctctcttaag
     3181 atctctttct ctcttgggtt atctgactct aaccagaatt gtgtcggtac gtgaaattgt
     3241 ttctctcttt ctgtttctat ttctaaaact aattttgtga aatgttgtac gtgagtgaca
     3301 aatacatctc tatgttgtct gtttatgatc ccatgtgaca tgtttaagat ctgttgctga
     3361 attggagatc tggatctgac ttctttttca tatgatccgc ttctgaattt tccatttgat
     3421 gttttttttt tgcagatttc aaggctcgag atggagatgg gtcgtctggt cttcgggttg
     3481 tgtttgattc ttattgtggg tacgtttctt gaattctccg gcggtgctac ggcgctgcag
     3541 cctggaggtg gaggtggagc tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3601 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3661 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3721 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc
     3781 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3841 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3901 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3961 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     4021 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     4081 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4141 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     4201 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4261 gacgagctgt acaaggatcc tgctggtgct gctgcgcctc tagcataatc tagagaattt
     4321 ccccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     4381 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     4441 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     4501 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     4561 atctatgtta ctgagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     4621 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4681 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4741 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4801 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4861 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4921 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgaaggc
     4981 cttgacagga tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt gagatctcat
     5041 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5101 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5161 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5221 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5281 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5341 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5401 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5461 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5521 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5581 cggaagaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5641 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5701 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5761 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5821 aatctaaagt actgatgaat cccctaatga ttttggtaaa aatcattaag ttaaggtgga
     5881 tacacatctt gtcatatgat caaatggttt cgcgaaaaat caataatcag acaacaagat
     5941 gtgcgaactc gatattttac acgactctct ttaccaattc tgccccgaat tacacttaaa
     6001 acgactcaac agcttaacgt tggcttgcca cgcattactt gactgtaaaa ctctcactct
     6061 taccgaactt ggccgtaacc tgccaaccaa agcgagaaca aaacataaca tcaaacgaat
     6121 cgaccgattg ttaggtaatc gtcacctcca caaagagcga ctcgctgtat accgttggca
     6181 tgctagcttt atctgttcgg gcaatacgat gcccattgta cttgttgact ggtctgatat
     6241 tcgtgagcaa aaacgactta tggtattgcg agcttcagtc gcactacacg gtcgttctgt
     6301 tactctttat gagaaagcgt tcccgctttc agagcaatgt tcaaagaaag ctcatgacca
     6361 atttctagcc gaccttgcga gcattctacc gagtaacacc acaccgctca ttgtcagtga
     6421 tgctggcttt aaagtgccat ggtataaatc cgttgagaag ctgggttggt actggttaag
     6481 tcgagtaaga ggaaaagtac aatatgcaga cctaggagcg gaaaactgga aacctatcag
     6541 caacttacat gatatgtcat ctagtcactc aaagacttta ggctataaga ggctgactaa
     6601 aagcaatcca atctcatgcc aaattctatt gtataaatct cgctctaaag gccgaaaaaa
     6661 tcagcgctcg acacggactc attgtcacca cccgtcacct aaaatctact cagcgtcggc
     6721 aaaggagcca tgggttctag caactaactt acctgttgaa attcgaacac ccaaacaact
     6781 tgttaatatc tattcgaagc gaatgcagat tgaagaaacc ttccgagact tgaaaagtcc
     6841 tgcctacgga ctaggcctac gccatagccg aacgagcagc tcagagcgtt ttgatatcat
     6901 gctgctaatc gccctgatgc ttcaactaac atgttggctt gcgggcgttc atgctcagaa
     6961 acaaggttgg gacaagcact tccaggctaa cacagtcaga aatcgaaacg tactctcaac
     7021 agttcgctta ggcatggaag ttttgcggca ttctggctac acaataacaa gggaagactt
     7081 actcgtggct gcaaccctac tagctcaaaa tttattcaca catggttacg ctttggggaa
     7141 attatgaggg gatctctcag tctaaagtat atatgtgtaa cattggtcta gtgattagaa
     7201 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     7261 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     7321 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     7381 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     7441 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     7501 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     7561 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     7621 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     7681 taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg
     7741 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     7801 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     7861 tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc
     7921 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga
     7981 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     8041 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     8101 ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
     8161 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
     8221 ctggtccacc tacaacaa
//
