LOCUS       sequence-420623-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning compatible vector for transient expression in 
            protoplasts. Arabidopsis UBQ10 promoter for N-terminal Dendra2 
            fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  AUTHORS   Allen JR, Flanagan M, Pathak S, Emenecker SK, Emenecker RJ, Strader 
            LC
  TITLE     An improved toolkit of gateway- and gibson assembly-compatible 
            vectors for protoplast transfection and agrobacterium-mediated plant
            transformation.
  JOURNAL   BMC Res Notes. 2025 Apr 8;18(1):149. doi: 
            10.1186/s13104-025-07231-1.
  PUBMED    40200324
REFERENCE   2  (bases 1 to 7496)
  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: "BMC Res 
            Notes."; date: "2025-04-8"; volume: "18(1):149. doi"; pages: " 
            10.1186/s13104-025-07231-1"
COMMENT     Sequence Label: pLCS105
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1989..2678
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     protein_bind    2748..2872
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2897..2902,2903..2920,2921..2927)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2981..3640
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3047..3066)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3982..4287
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4166..4185
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4328..4452)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4511..5218
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(5275..5291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5275..5291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5288..5310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5299..5315
                     /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)."
     promoter        complement(join(5323..5329,5330..5347,5348..5353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5368..5389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5506..5523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5677..6265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5757..5776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6436..7227,7228..7296))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7059..7078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7297..7401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7469..7487
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa actttcgcga agcacccacc
      421 ctaagccaca ttttaatcct tctgttggcc atgccccatc aaagttgcac ttaacccaag
      481 attgtggtgg agcttcccat gtttctcgtc tgtcccgacg gtgttgtggt tggtgctttc
      541 cttacattct gagcctcttt ccttctaatc cactcatctg catcttcttg tgtccttact
      601 aatacctcat tggttccaaa ttccctccct ttaagcacca gctcgtttct gttcttccac
      661 agcctcccaa gtatccaagg gactaaagcc tccacattct tcagatcagg atattcttgt
      721 ttaagatgtt gaactctatg gaggtttgta tgaactgatg atctaggacc ggataagttc
      781 ccttcttcat agcgaactta ttcaaagaat gttttgtgta tcattcttgt tacattgtta
      841 ttaatgaaaa aatattattg gtcattggac tgaacacgag tgttaaatat ggaccaggcc
      901 ccaaataaga tccattgata tatgaattaa ataacaagaa taaatcgagt caccaaacca
      961 cttgcctttt ttaacgagac ttgttcacca acttgataca aaagtcatta tcctatgcaa
     1021 atcaataatc atacaaaaat atccaataac actaaaaaat taaaagaaat ggataatttc
     1081 acaatatgtt atacgataaa gaagttactt ttccaagaaa ttcactgatt ttataagccc
     1141 acttgcatta gataaatggc aaaaaaaaac aaaaaggaaa agaaataaag cacgaagaat
     1201 tctagaaaat acgaaatacg cttcaatgca gtgggaccca cggttcaatt attgccaatt
     1261 ttcagctcca ccgtatattt aaaaaataaa acgataatgc taaaaaaata taaatcgtaa
     1321 cgatcgttaa atctcaacgg ctggatctta tgacgaccgt tagaaattgt ggttgtcgac
     1381 gagtcagtaa taaacggcgt caaagtggtt gcagccggca cacacgagtc gtgtttatca
     1441 actcaaagca caaatacttt tcctcaacct aaaaataagg caattagcca aaaacaactt
     1501 tgcgtgtaaa caacgctcaa tacacgtgtc attttattat tagctattgc ttcaccgcct
     1561 tagctttctc gtgacctagt cgtcctcgtc ttttcttctt cttcttctat aaaacaatac
     1621 ccaaagagct cttcttcttc acaattcaga tttcaatttc tcaaaatctt aaaaactttc
     1681 tctcaattct ctctaccgtg atcaaggtaa atttctgtgt tccttattct ctcaaaatct
     1741 tcgattttgt tttcgttcga tcccaatttc gtatatgttc tttggtttag attctgttaa
     1801 tcttagatcg aagacgattt tctgggtttg atcgttagat atcatcttaa ttctcgatta
     1861 gggtttcata gatatcatcc gatttgttca aataatttga gttttgtcga ataattactc
     1921 ttcgatttgt gatttctatc tagatctggt gttagtttct agtttgtgcg atcgaatttg
     1981 tcctcgagat gaacaccccg ggaattaacc tgatcaagga ggacatgcgc gtgaaggtgc
     2041 acatggaggg caacgtgaac ggccacgcct tcgtgatcga gggcgagggc aagggcaagc
     2101 cctacgaggg cacccagacc gccaacctga ccgtgaagga gggcgccccc ctgcccttca
     2161 gctacgacat cctgaccacc gccgtgcact acggcaaccg ggtgttcacc aagtaccccg
     2221 aggacatccc cgactacttc aagcagagct tccccgaggg ctacagctgg gagcgcacca
     2281 tgaccttcga ggacaagggc atctgcacca tccgcagcga catcagcctg gagggcgact
     2341 gcttcttcca gaacgtgcgc ttcaagggca ccaacttccc ccccaacggc cccgtgatgc
     2401 agaagaagac cctgaagtgg gagcccagca ccgagaagct gcacgtgcgc gacggcctgc
     2461 tggtgggcaa catcaacatg gccctgctgc tggagggcgg cggccactac ctgtgcgact
     2521 tcaagaccac ctacaaggcc aagaaggtgg tgcagctgcc cgacgcccac ttcgtggacc
     2581 accgcatcga gatcctgggc aacgacagcg actacaacaa ggtgaagctg tacgagcacg
     2641 ccgtggcccg ctacagcccc ctgcccagcc aggtgtggtc cggactcaga tcacgagctc
     2701 aagcttcaaa ttctgcagtc gacggtaccg cgggcccggg atcatcaaca agtttgtaca
     2761 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     2821 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc
     2881 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
     2941 ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     3001 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     3061 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     3121 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     3181 gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     3241 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     3301 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     3361 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     3421 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     3481 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     3541 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     3601 attacaacag tactgcgatg agtggcaggg cggggcgtaa agatctggat ccggcttact
     3661 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     3721 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     3781 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     3841 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     3901 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     3961 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     4021 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     4081 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     4141 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     4201 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     4261 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     4321 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     4381 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     4441 gtacaaagtg gttgatgatc ctaggtgagt ctagagagtt aattaagacc cgggactagt
     4501 ccctagagtc ctgctttaat gagatatgcg agacgcctat gatcgcatga tatttgcttt
     4561 caattctgtt gtgcacgttg taaaaaacct gagcatgtgt agctcagatc cttaccgccg
     4621 gtttcggttc attctaatga atatatcacc cgttactatc gtatttttat gaataatatt
     4681 ctccgttcaa tttactgatt gtaccctact acttatatgt acaatattaa aatgaaaaca
     4741 atatattgtg ctgaataggt ttatagcgac atctatgata gagcgccaca ataacaaaca
     4801 attgcgtttt attattacaa atccaatttt aaaaaaagcg gcagaaccgg tcaaacctaa
     4861 aagactgatt acataaatct tattcaaatt tcaaaagtgc cccaggggct agtatctacg
     4921 acacaccgag cggcgaacta ataacgctca ctgaagggaa ctccggttcc ccgccggcgc
     4981 gcatgggtga gattccttga agttgagtat tggccgtccg ctctaccgaa agttacgggc
     5041 accattcaac ccggtccagc acggcggccg ggtaaccgac ttgctgcccc gagaattatg
     5101 cagcattttt ttggtgtatg tgggccccaa atgaagtgca ggtcaaacct tgacagtgac
     5161 gacaaatcgt tgggcgggtc cagggcgaat tttgcgacaa catgtcgagg ctcagcagga
     5221 cctgcaggtc gactctagag gatccccggg taccgagctc gaattcgtaa tcatgtcata
     5281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5341 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7441 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
