LOCUS       sequence-360342-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant transient expression vector containing eCFP tag. Unique 
            nucleotide sequences Uc and Ud for cloning at N terminus of eCFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  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 4732)
  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/N-eCFP
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        536..880
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     823..843
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    898..922
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             934..954
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1051..1053,1054..1056,1057..1569)
                     /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(1096..1117)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1357..1376)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1704..1725
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1772..2023
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2080..2097)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2080..2096)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2089..2111)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2305..2324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2424..2446
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2484..2502)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2570..2674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2675..2743,2744..3535)
                     /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(2893..2912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3706..4294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4195..4214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4448..4465
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4582..4603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4618..4623,4624..4641,4642..4648)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4656..4672
                     /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     4661..4683
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4680..4696
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4680..4696
                     /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 aagaagacgt tccaaccacg tcttcaaagc aagtggattg
      781 atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc
      841 ttcctctata taaggaagtt catttcattt ggagagaaca cgggggactc tagatcaaca
      901 agtttgtaca aaaaagcagg ctccgcggcc gccttgttta actttaagaa ggagcccttc
      961 accagccaac tccctttaca acctcactca agtccgttag agcccgggct cgttcgctgc
     1021 cacctaagaa tactctacgg tcacatacct atggtgagca agggcgagga gctgttcacc
     1081 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1141 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1201 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag
     1261 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1321 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1381 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1441 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat cagcgacaac
     1501 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1561 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1621 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     1681 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgcctcggct
     1741 tagtaataac tcgagaagct cgaatttccc cgatcgttca aacatttggc aataaagttt
     1801 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
     1861 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggttttatg
     1921 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
     1981 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcggggttc cttatcatct
     2041 ggcgaatcgg acccacaaga gcactgggcg cgccaattca ctggccgtcg ttttacaacg
     2101 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2161 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2221 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2281 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2341 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2401 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2461 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2521 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2581 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2641 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2701 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2761 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2821 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2881 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2941 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3001 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3061 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3121 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3181 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3241 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3301 ggaggcggat aaagttgcag gaccacttct gcgctcggtc cttccggctg gctggtttat
     3361 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3421 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3481 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3541 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3601 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3661 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3721 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3781 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3841 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3901 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3961 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4021 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4081 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4141 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4201 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4261 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgct agcaacgcgg cctttttacg
     4321 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4381 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4441 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     4501 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4561 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     4621 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4681 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcc cc
//
