LOCUS       sequence-361430-        4736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant transient expression vector containing eCFP tag. Unique 
            nucleotide sequences Uc and Ud for cloning at C terminus of eCFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4736)
  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 4736)
  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: pUC19/eCFP-C
FEATURES             Location/Qualifiers
     source          1..4736
                     /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             join(965..967,968..970,971..1483)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1010..1031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1271..1290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1618..1639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1776..2027
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2084..2101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2084..2100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2309..2328)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2428..2450
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2488..2506)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2574..2678
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2679..2747,2748..3539)
                     /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(2897..2916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3710..4298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4199..4218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4452..4469
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4586..4607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4622..4627,4628..4645,4646..4652)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4660..4676
                     /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     4665..4687
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4684..4700
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4684..4700
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1021 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1141 caccctcgtg accaccctga cctggggcgt gcagtgcttc gcccgctacc ccgaccacat
     1201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1381 gcacaagctg gagtacaacg ccatcagcga caacgtctat atcaccgccg acaagcagaa
     1441 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1561 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1621 ggtcctgctg gagttcgtga ccgccgcctc ggctcagagc caactccctt tacaacctca
     1681 ctcaagtccg ttagagcccg ggctcgttcg ctgccaccta agaatactct acggtcacat
     1741 accctgataa taactcgaga agctcgaatt tccccgatcg ttcaaacatt tggcaataaa
     1801 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     1861 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     1921 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc
     1981 aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg gttccttatc
     2041 atctggcgaa tcggacccac aagagcactg ggcgcgctaa ttcactggcc gtcgttttac
     2101 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2161 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2221 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2281 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2341 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2401 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2461 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2521 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2581 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2641 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2701 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2761 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2821 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2881 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2941 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3001 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3061 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3241 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggtccttccg gctggctggt
     3361 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3421 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3541 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3601 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3661 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4081 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4141 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4201 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4261 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgctagcaac gcggcctttt
     4321 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4381 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4441 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4501 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4561 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4621 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4681 acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag gtcccc
//
