LOCUS       sequence-360385-        5249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of Golgi apparatus-targeted 
            mCherry in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5249)
  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 5249)
  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/mChe-Golgi
FEATURES             Location/Qualifiers
     source          1..5249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        535..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             964..1671
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1109..1127)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1377..1396
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2288..2540
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2597..2614)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2597..2613)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2606..2628)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2822..2841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2941..2963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3001..3019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3087..3191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3192..3260,3261..4052)
                     /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(3410..3429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4223..4811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4712..4731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4965..4982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5099..5120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5135..5140,5141..5158,5159..5165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5173..5189
                     /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     5178..5200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5197..5213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5197..5213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccat tactcgcatc cattctcagg ctgtctcgtc tcgtctcaga ttagcctttt
       61 caatttcaga aagaatgcta acccacagat ggttagagag gcttacgcag caggtctcat
      121 caagacgatc tacccgagca ataatctcca ggaaatcaaa taccttccca agaaggttaa
      181 agatgcagtc aaaagattca ggactaactg catcaagaac acagagaaag atatatttct
      241 caagatcaga agtactattc cagtatggac gattcaaggc ttgcttcaca aaccaaggca
      301 agtaatagag attggagtct ctaaaaaggt agttcccact gaatcaaagg ccatggagtc
      361 aaagattcaa atagaggacc taacagaact cgccgtaaag actggcgaac agttcataca
      421 gagtctctta cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac
      481 acttgtctac tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac
      541 ttttcaacaa agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca
      601 ctttattgtg aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa
      661 aggaaaggcc atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
      721 cacgaggagc atcgtggaaa 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 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1021 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1081 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1141 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1201 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1261 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1321 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1381 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1441 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1501 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1561 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1621 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gatggaagga
     1681 agcggtggtg acagtggttc gatggctacg cctgtacaaa agaaagtaca tgaggcatgg
     1741 agggtttacc aatattatct ggacaagact acacctcatt caactaacag gtggatcgga
     1801 acacttgttt tctttcttat ctactgtttg agggtttact ctattcacgg attctacatc
     1861 atctcctatg gtcttggaat ctacctcctg aatctgctta tcgggttttt gtctcctctt
     1921 gttgatcctg agctcgaggt atctgatgga gctactctgc ctactcgagg ttctgatgag
     1981 ttcaagccat ttatccgccg tctcccggag tttaaattct ggtactccat gactaaggca
     2041 ttctgcatag cctttcttat gacattcttc tcggtgtttg atgttcccgt cttctggccc
     2101 atactgcttt gctactgggt cgttctcttt gtcctcacca tgagacgcca aatagcccac
     2161 atgatcaagc acaagtacat tcctttcagc atcggaaaac agaaatatag cggccggaaa
     2221 tcttctgcga acagcggcgg cggctctcgt gctgattgat aataactcga gaagctcgaa
     2281 tttccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     2341 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     2401 gtaatgcatg acgttattta tgagatgggt tttatgatta gagtcccgca attatacatt
     2461 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     2521 tcatctaatg ttactagatc gggccaggat acatagatta ccacaactcc gagcccttcc
     2581 accggcgcgc caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2641 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2701 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     2761 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     2821 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     2881 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2941 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3001 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3061 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3121 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3181 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3241 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3301 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3361 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3421 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3481 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3541 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3601 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3661 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3721 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3781 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3841 cacttctgcg ctcggtcctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3901 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3961 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4021 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4081 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4801 gcctatggaa aaacgctagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4861 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4921 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4981 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5041 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5101 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5161 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5221 acgccaagct tgcatgcctg caggtcccc
//
