LOCUS       sequence-360169-        4829 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 N terminus of Vn..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4829)
  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 4829)
  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-Vn
FEATURES             Location/Qualifiers
     source          1..4829
                     /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..1547)
                     /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(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     complement(1965..1984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1966..1984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2105..2764)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2679..2698
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3136..3720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3621..3640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3874..3891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3906..4046
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3972..3991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4132..4154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    4292..4313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4328..4333,4334..4351,4352..4358)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4366..4382
                     /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     4371..4393
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4390..4406
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4390..4406
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4402..4644
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPDIMSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE
                     ARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRS"
     primer_bind     complement(4431..4448)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4431..4447)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4440..4462)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4651..4668
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(4681..4714)
                     /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 aagctgatct gcaccaccgg caagctgccc gtgccctggc
     1261 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac 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 tatcacctaa tcacctgatc
     1561 tagagtccgc aaaaatcacc agtctctctc tacaaatcta tctctctcta tttttctcca
     1621 gaataatgtg tgagtagttc ccagataagg gaattagggt tcttataggg tttcgctcat
     1681 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
     1741 aaaatttcta attcctaaaa ccaaaatcca gtgacgctgg gagttcgtag acggaaacaa
     1801 acgcagaatc caagcctgca gggttcctta tcatctggcg aatcggaccc acaagagcac
     1861 tggtttaaac gcggccgcat aaggccggcc ttgccctagt tcctatagcc tacattatac
     1921 gaagttatac tagggataac agggtaattc agatctcagc gcgcccctat agtgagtcgt
     1981 attacgaatg ggagcaatat tgtgcttagt gcatctaacg cttgagttaa gccgcgccgc
     2041 gaagcggcgt cggcttgaac gaatttctag ctagaggatc gcaccaataa ctgccttaaa
     2101 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     2161 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     2221 tgtcgccttg cgtataatat ttgcccattg tgaaaacggg ggcgaagaag ttgtccatat
     2281 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     2341 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     2401 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     2461 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2521 gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa
     2581 tgtgaataaa ggccggataa aacttgtgct tacttttctt tacggtcttt aaaaaggccg
     2641 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     2701 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     2761 ccatgatgtt taactttgtt ttagggcgac tgccctgctg cgtaatattg aaggttctcg
     2821 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2881 gacgggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2941 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3001 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctacgcgtg
     3061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3181 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3241 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3301 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3361 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3421 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3481 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3541 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3601 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3661 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3721 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3781 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3841 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3901 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3961 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     4021 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gacgcgtcgc ccaatacgca
     4201 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4261 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4321 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4381 aatttcacac aggaaacagc tatgaccatg attacgccag atatcatgtc actggccgtc
     4441 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4501 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4561 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4621 ggtgtggtgg ttacgcgcag ctaagatatc atttaggtga cactatagcc aagcgcgctc
     4681 ataacttcgt atagcataca ttatacgaag ttatcagatc taatactgcc atttcattac
     4741 ctctttctcc gcacccgaca tagatgcaat aacttcgtat aggctaattt atacgatgta
     4801 ggagatcgat gcattgcggc cgctagctc
//
