LOCUS       sequence-360085-        5087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant transient expression vector containing eGFP tag. Unique 
            nucleotide sequences Uc and Ud for cloning at N terminus of eGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5087)
  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 5087)
  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/N-eGFP
FEATURES             Location/Qualifiers
     source          1..5087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        511..855
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(1086..1088,1089..1091,1092..1805)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1086..1088,1089..1091,1092..1802)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1131..1152)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1392..1411)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1739..1760
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2223..2242)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2224..2242)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2363..3022)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2937..2956
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3394..3978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3879..3898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4132..4149
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4164..4304
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4230..4249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4390..4412
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    4550..4571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4586..4591,4592..4609,4610..4616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4624..4640
                     /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     4629..4651
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4648..4664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4648..4664
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4660..4902
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPDIMSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE
                     ARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRS"
     primer_bind     complement(4689..4706)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4689..4705)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4698..4720)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4909..4926
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(4939..4972)
                     /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 ggacccttcc tctatataag gaagttcatt
      841 tcatttggag aggacgtcga gagttctcaa cacaacatat acaaaacaaa cgaatctcaa
      901 gcaatcaagc attctacttc tattgcagca atttaaatca tttcttttaa agcaaaagca
      961 attttctgaa aattttcacc atttacgaac gatagccgag ccaactccct ttacaacctc
     1021 actcaagtcc gttagagccc gggctcgttc gctgccacct aagaatactc tacggtcaca
     1081 tacctatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1141 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1201 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1261 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1321 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1381 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1441 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1501 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1561 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1621 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1681 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1741 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1801 agtaataatc acctgatcta gagtccgcaa aaatcaccag tctctctcta caaatctatc
     1861 tctctctatt tttctccaga ataatgtgtg agtagttccc agataaggga attagggttc
     1921 ttatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
     1981 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt gacgctggga
     2041 gttcgtagac ggaaacaaac gcagaatcca agcctgcagg gttccttatc atctggcgaa
     2101 tcggacccac aagagcactg gtttaaacgc ggccgcataa ggccggcctt gccctagttc
     2161 ctatagccta cattatacga agttatacta gggataacag ggtaattcag atctcagcgc
     2221 gcccctatag tgagtcgtat tacgaatggg agcaatattg tgcttagtgc atctaacgct
     2281 tgagttaagc cgcgccgcga agcggcgtcg gcttgaacga atttctagct agaggatcgc
     2341 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
     2401 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2461 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccattgtg aaaacggggg
     2521 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2581 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2641 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2701 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2761 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat
     2821 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta cttttcttta
     2881 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     2941 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     3001 atccagtgat ttttttctcc atgatgttta actttgtttt agggcgactg ccctgctgcg
     3061 taatattgaa ggttctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3121 tatcgtagtt atctacacga cgggagtcag gcaactatgg atgaacgaaa tagacagatc
     3181 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3241 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3301 tttgataatc tacgcgtgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3361 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3421 ctgctgcttg caaacaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4081 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4141 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4201 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4261 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     4321 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4381 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4441 cgcgtcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4501 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4561 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4621 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat
     4681 atcatgtcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4741 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4801 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     4861 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagct aagatatcat ttaggtgaca
     4921 ctatagccaa gcgcgctcat aacttcgtat agcatacatt atacgaagtt atcagatcta
     4981 atactgccat ttcattacct ctttctccgc acccgacata gatgcaataa cttcgtatag
     5041 gctaatttat acgatgtagg agatcgatgc attgcggccg ctagctc
//
