LOCUS       sequence-360159-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bimolecular fluorescence complementation (BiFC) vector for transient
            expression of N-truncated Venus in plants. Unique nucleotide 
            sequences Uc and Ud for cloning at C terminus of Vn..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  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 4835)
  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/Vn-C
FEATURES             Location/Qualifiers
     source          1..4835
                     /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..1471)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIT"
     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     complement(1971..1990)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1972..1990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2111..2770)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2685..2704
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3142..3726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3627..3646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3880..3897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3912..4052
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3978..3997)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4138..4160
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    4298..4319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4334..4339,4340..4357,4358..4364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4372..4388
                     /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     4377..4399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4396..4412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4396..4412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4408..4650
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPDIMSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE
                     ARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRS"
     primer_bind     complement(4437..4454)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4437..4453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4446..4468)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4657..4674
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(4687..4720)
                     /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 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1201 cctgggctac ggcctgcagt gcttcgcccg 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 tctatatcac ccagagccaa ctccctttac aacctcactc
     1501 aagtccgtta gagcccgggc tcgttcgctg ccacctaaga atactctacg gtcacatacc
     1561 ctgatctaga gtccgcaaaa atcaccagtc tctctctaca aatctatctc tctctatttt
     1621 tctccagaat aatgtgtgag tagttcccag ataagggaat tagggttctt atagggtttc
     1681 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
     1741 atcaataaaa tttctaattc ctaaaaccaa aatccagtga cgctgggagt tcgtagacgg
     1801 aaacaaacgc agaatccaag cctgcagggt tccttatcat ctggcgaatc ggacccacaa
     1861 gagcactggt ttaaacgcgg ccgcataagg ccggccttgc cctagttcct atagcctaca
     1921 ttatacgaag ttatactagg gataacaggg taattcagat ctcagcgcgc ccctatagtg
     1981 agtcgtatta cgaatgggag caatattgtg cttagtgcat ctaacgcttg agttaagccg
     2041 cgccgcgaag cggcgtcggc ttgaacgaat ttctagctag aggatcgcac caataactgc
     2101 cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     2161 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     2221 gcaccttgtc gccttgcgta taatatttgc ccattgtgaa aacgggggcg aagaagttgt
     2281 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     2341 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     2401 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     2461 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     2521 tcaccagctc accgtctttc attgccatac ggaactccgg atgagcattc atcaggcggg
     2581 caagaatgtg aataaaggcc ggataaaact tgtgcttact tttctttacg gtctttaaaa
     2641 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     2701 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     2761 ttttctccat gatgtttaac tttgttttag ggcgactgcc ctgctgcgta atattgaagg
     2821 ttctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2881 ctacacgacg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatcta
     3061 cgcgtgcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3121 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3181 aacaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3781 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3841 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3901 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3961 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     4021 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     4081 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4141 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgacg cgtcgcccaa
     4201 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4261 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4321 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4381 gataacaatt tcacacagga aacagctatg accatgatta cgccagatat catgtcactg
     4441 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4501 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4561 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     4621 gcggcgggtg tggtggttac gcgcagctaa gatatcattt aggtgacact atagccaagc
     4681 gcgctcataa cttcgtatag catacattat acgaagttat cagatctaat actgccattt
     4741 cattacctct ttctccgcac ccgacataga tgcaataact tcgtataggc taatttatac
     4801 gatgtaggag atcgatgcat tgcggccgct agctc
//
