LOCUS       sequence-360087-        5090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant transient expression vector containing eGFP tag. Unique 
            nucleotide sequences Uc and Ud for cloning at C terminus of eGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5090)
  AUTHORS   Han J, Ma K, Li H, Su J, Zhou L, Tang J, Zhang S, Hou Y, Chen L, Liu
            YG, Zhu Q
  TITLE     All-in-one: A Robust Fluorescent Fusion Protein Vector Toolbox for 
            Protein Localization and BiFC Analyses in Plants.
  JOURNAL   Plant Biotechnol J. 2022 Feb 18. doi: 10.1111/pbi.13790.
  PUBMED    35179286
REFERENCE   2  (bases 1 to 5090)
  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 
            Biotechnol J. 2022 Feb 18. doi: 10.1111/pbi.13790."
COMMENT     Sequence Label: pYL322d1/eGFP-C
FEATURES             Location/Qualifiers
     source          1..5090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        511..855
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     799..819
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(1010..1012,1013..1015,1016..1726)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1010..1012,1013..1015,1016..1726)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1055..1076)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1316..1335)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1663..1684
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2226..2245)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2227..2245)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2366..3025)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2940..2959
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3397..3981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3882..3901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4135..4152
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4233..4252)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4393..4415
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    4553..4574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4589..4594,4595..4612,4613..4619)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4627..4643
                     /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)."
     primer_bind     4632..4654
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4651..4667
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4651..4667
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4663..4905
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPDIMSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE
                     ARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRS"
     primer_bind     complement(4692..4709)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4692..4708)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4701..4723)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4912..4929
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(4942..4975)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gtttaaacca ttactcgcat ccattctcag gctgtctcgt ctcgtctcgg cgcgccccag
       61 gatacataga ttaccacaac tccgagccct tccaccgcgc cttaccggtc aacatgtgga
      121 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
      181 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
      241 tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca
      301 ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg
      361 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
      421 agtggattga tgtgataaca tggtggagca cgacacactt gtctactcca aaaatatcaa
      481 agatacagtc tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg
      541 aaacctcctc ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa
      601 ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc
      661 ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga
      721 agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag
      781 ggatgacgca caatcccact atccttcgca agacccttcc tctatataag gaagttcatt
      841 tcatttggag aggacgtcga gagttctcaa cacaacatat acaaaacaaa cgaatctcaa
      901 gcaatcaagc attctacttc tattgcagca atttaaatca tttcttttaa agcaaaagca
      961 attttctgaa aattttcacc atttacgaac gatagccatg ggcactagta tggtgagcaa
     1021 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1081 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1141 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1201 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1261 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1321 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1381 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1441 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1501 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1561 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1621 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1681 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagcaga gccaactccc
     1741 tttacaacct cactcaagtc cgttagagcc cgggctcgtt cgctgccacc taagaatact
     1801 ctacggtcac ataccctgat ctagagtccg caaaaatcac cagtctctct ctacaaatct
     1861 atctctctct atttttctcc agaataatgt gtgagtagtt cccagataag ggaattaggg
     1921 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     1981 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgacgctg
     2041 ggagttcgta gacggaaaca aacgcagaat ccaagcctgc agggttcctt atcatctggc
     2101 gaatcggacc cacaagagca ctggtttaaa cgcggccgca taaggccggc cttgccctag
     2161 ttcctatagc ctacattata cgaagttata ctagggataa cagggtaatt cagatctcag
     2221 cgcgccccta tagtgagtcg tattacgaat gggagcaata ttgtgcttag tgcatctaac
     2281 gcttgagtta agccgcgccg cgaagcggcg tcggcttgaa cgaatttcta gctagaggat
     2341 cgcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
     2401 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     2461 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatt gtgaaaacgg
     2521 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     2581 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     2641 caccgtaaca cgccacatct tgcgaatata tgtgtaaaaa ctgccggaaa tcgtcgtggt
     2701 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     2761 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaay tccggatgag
     2821 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttacttttct
     2881 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     2941 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     3001 tatatccagt gatttttttc tccatgatgt ttaactttgt tttagggcga ctgccctgct
     3061 gcgtaatatt gaaggttctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3121 ccgtatcgta gttatctaca cgacgggagt caggcaacta tggatgaacg aaatagacag
     3181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3301 ctttttgata atctacgcgt gcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3421 aatctgctgc ttgcaaacaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3481 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3541 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3601 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3661 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3721 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3781 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3841 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3901 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3961 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4021 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4081 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4141 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     4201 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4261 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgygccc cgacacccgc
     4321 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4381 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4441 cgacgcgtcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4501 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     4561 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     4621 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     4681 gatatcatgt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4741 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4801 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4861 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gctaagatat catttaggtg
     4921 acactatagc caagcgcgct cataacttcg tatagcatac attatacgaa gttatcagat
     4981 ctaatactgc catttcatta cctctttctc cgcacccgac atagatgcaa taacttcgta
     5041 taggctaatt tatacgatgt aggagatcga tgcattgcgg ccgctagctc
//
