LOCUS       sequence-356728-        8217 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Basic chitinase 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 8217)
  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 8217)
  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 
            Direct."; date: "2021-11-28"; volume: "5(11):e360. doi"; pages: " 
            10.1002/pld3.360. eCollection 2021 Nov"
COMMENT     Sequence Label: secCitrine
FEATURES             Location/Qualifiers
     source          1..8217
                     /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(3541..3543,3544..4254)
                     /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(3583..3604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3844..3863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4191..4212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4570..4590)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4570..4588)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4609..4625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4609..4625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4622..4644)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4633..4649
                     /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(4657..4663,4664..4681,4682..4687))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4702..4723
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4840..4857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4962..4986
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5077..5665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5157..5176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7174..7989)
                     /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(7174..7989)
                     /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     7897..7916
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7981..8000
                     /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 atgaagacta atctttttct ctttctcatc
     3481 ttttcacttc tcctatcatt atcctcggcc gagctgcagc ctggaggtgg aggtggagct
     3541 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3601 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     3661 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3721 gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag
     3781 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3841 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3901 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3961 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4021 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4081 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4141 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4201 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggatcct
     4261 gctggtgctg ctgcgcctct agcataatct agagaatttc cccgatcgtt caaacatttg
     4321 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     4381 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     4441 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     4501 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tgagctccag
     4561 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     4621 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4681 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4741 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4801 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4861 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4921 cacagaatca ggggataacg caggaaagaa catgaaggcc ttgacaggat atattggcgg
     4981 gtaaactaag tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa
     5041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaagaagag ttggtagctc
     5581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta ctgatgaatc
     5821 ccctaatgat tttggtaaaa atcattaagt taaggtggat acacatcttg tcatatgatc
     5881 aaatggtttc gcgaaaaatc aataatcaga caacaagatg tgcgaactcg atattttaca
     5941 cgactctctt taccaattct gccccgaatt acacttaaaa cgactcaaca gcttaacgtt
     6001 ggcttgccac gcattacttg actgtaaaac tctcactctt accgaacttg gccgtaacct
     6061 gccaaccaaa gcgagaacaa aacataacat caaacgaatc gaccgattgt taggtaatcg
     6121 tcacctccac aaagagcgac tcgctgtata ccgttggcat gctagcttta tctgttcggg
     6181 caatacgatg cccattgtac ttgttgactg gtctgatatt cgtgagcaaa aacgacttat
     6241 ggtattgcga gcttcagtcg cactacacgg tcgttctgtt actctttatg agaaagcgtt
     6301 cccgctttca gagcaatgtt caaagaaagc tcatgaccaa tttctagccg accttgcgag
     6361 cattctaccg agtaacacca caccgctcat tgtcagtgat gctggcttta aagtgccatg
     6421 gtataaatcc gttgagaagc tgggttggta ctggttaagt cgagtaagag gaaaagtaca
     6481 atatgcagac ctaggagcgg aaaactggaa acctatcagc aacttacatg atatgtcatc
     6541 tagtcactca aagactttag gctataagag gctgactaaa agcaatccaa tctcatgcca
     6601 aattctattg tataaatctc gctctaaagg ccgaaaaaat cagcgctcga cacggactca
     6661 ttgtcaccac ccgtcaccta aaatctactc agcgtcggca aaggagccat gggttctagc
     6721 aactaactta cctgttgaaa ttcgaacacc caaacaactt gttaatatct attcgaagcg
     6781 aatgcagatt gaagaaacct tccgagactt gaaaagtcct gcctacggac taggcctacg
     6841 ccatagccga acgagcagct cagagcgttt tgatatcatg ctgctaatcg ccctgatgct
     6901 tcaactaaca tgttggcttg cgggcgttca tgctcagaaa caaggttggg acaagcactt
     6961 ccaggctaac acagtcagaa atcgaaacgt actctcaaca gttcgcttag gcatggaagt
     7021 tttgcggcat tctggctaca caataacaag ggaagactta ctcgtggctg caaccctact
     7081 agctcaaaat ttattcacac atggttacgc tttggggaaa ttatgagggg atctctcagt
     7141 ctaaagtata tatgtgtaac attggtctag tgattagaaa aactcatcga gcatcaaatg
     7201 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     7261 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     7321 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     7381 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     7441 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     7501 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     7561 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     7621 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     7681 ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     7741 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     7801 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     7861 caatcggtag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     7921 taaatcagca tccatgttgg aatttaatcg cggccttgag caagacgttt cccgttgaat
     7981 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     8041 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg
     8101 ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc
     8161 agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaa
//
