LOCUS       sequence-360348-        4441 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bimolecular fluorescence complementation (BiFC) vector for transient
            expression of C-truncated Venus in plants. Unique nucleotide 
            sequences Uc and Ud for cloning at C terminus of Vc..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4441)
  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 4441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pUC19/Vc-C
FEATURES             Location/Qualifiers
     source          1..4441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        536..881
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     824..844
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    899..923
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             935..955
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             971..997
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1022..1273
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1210..1231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1395..1646
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1789..1805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1798..1820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2014..2033)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2133..2155
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2193..2211)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2279..2383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2384..2452,2453..3244)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSV
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2602..2621)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3415..4003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3904..3923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4157..4174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4291..4312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4327..4332,4333..4350,4351..4357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4365..4381
                     /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     4370..4392
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4389..4405
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4389..4405
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccgc tgggagttcg tagacggaaa caaacgcaga atccaagcag attagccttt
       61 tcaatttcag aaagaatgct aacccacaga tggttagaga ggcttacgca gcaggtctca
      121 tcaagacgat ctacccgagc aataatctcc aggaaatcaa ataccttccc aagaaggtta
      181 aagatgcagt caaaagattc aggactaact gcatcaagaa cacagagaaa gatatatttc
      241 tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcac aaaccaaggc
      301 aagtaataga gattggagtc tctaaaaagg tagttcccac tgaatcaaag gccatggagt
      361 caaagattca aatagaggac ctaacagaac tcgccgtaaa gactggcgaa cagttcatac
      421 agagtctctt acgactcaat gacaagaaga aaatcttcgt caacatggtg gagcacgaca
      481 cacttgtcta ctccaaaaat atcaaagata cagtctcaga agaccaaagg gcaattgaga
      541 cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca gctatctgtc
      601 actttattgt gaagatagtg gaaaaggaag gtggctccta caaatgccat cattgcgata
      661 aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac
      721 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
      781 gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
      841 cttcctctat ataaggaagt tcatttcatt tggagagaac acgggggact ctagatcaac
      901 aagtttgtac aaaaaagcag gctccgcggc cgccttgttt aactttaaga aggagccctt
      961 caccatggag tacccatacg acgtaccaga ttacgctcat atggccatga tcaagcttgc
     1021 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     1081 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1141 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
     1201 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1261 cgagctgtac aagcagagcc aactcccttt acaacctcac tcaagtccgt tagagcccgg
     1321 gctcgttcgc tgccacctaa gaatactcta cggtcacata ccctgataat aactcgagaa
     1381 gctcgaattt ccccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg
     1441 ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa
     1501 ttaacatgta atgcatgacg ttatttatga gatgggtttt atgattagag tcccgcaatt
     1561 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     1621 cgcggtgtca tctatgttac tagatcgggg ttccttatca tctggcgaat cggacccaca
     1681 agagcactgg gcgcgccaat tcactggccg tcgtcatcta tgttactaga tcggtgttcc
     1741 ttatcatctg gcgaatcgga cccacaagag cactgggcgc gccaattcac tggccgtcgt
     1801 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1861 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1921 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     1981 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2041 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2101 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2161 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     2221 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2281 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2341 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2401 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2461 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2521 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2581 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2641 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2701 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2761 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2821 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2881 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2941 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3001 agactggatg gaggcggata aagttgcagg accacttctg cgctcggtcc ttccggctgg
     3061 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3121 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3181 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3241 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3301 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3361 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3541 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcta gcaacgcggc
     4021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4081 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4141 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4201 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     4261 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     4321 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     4381 atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtccc
     4441 c
//
