LOCUS       sequence-420447-        8246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning compatible vector for transient expression in 
            protoplasts. Arabidopsis UBQ10 promoter expression of N-terminal 
            mScarlet-I-mNeonGreen tandem fluorescent timer fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  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 8246)
  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: pLCS111
FEATURES             Location/Qualifiers
     source          1..8246
                     /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"
     protein_bind    3498..3622
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3647..3652,3653..3670,3671..3677)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3731..4390
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3797..3816)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4732..5037
                     /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     4916..4935
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5078..5202)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      5261..5968
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(6025..6041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6025..6041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6038..6060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6049..6065
                     /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(6073..6079,6080..6097,6098..6103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6118..6139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6256..6273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6427..7015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6507..6526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7186..7977,7978..8046))
                     /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     7809..7828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8047..8151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8219..8237
                     /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 ggtcagtaaa ggcgaggcag tgatcaaaga attcatgcgt ttcaaggtcc
     2041 acatggaagg gtcaatgaac ggtcacgaat ttgagataga aggggaaggc gagggacgac
     2101 cgtacgaggg tacccagacc gcaaaactta aagtcaccaa gggtgggcca ttgcctttct
     2161 cttgggacat actatcccct cagtttatgt atggttcgag ggcatttatc aaacacccag
     2221 cggacattcc agactattat aaacagtctt tccctgaagg attcaaatgg gaacgagtca
     2281 tgaacttcga ggacggtggc gcggtaacag tcacgcagga cacctcgttg gaagacggca
     2341 cgctcatata taaagtgaaa ctacggggta ctaacttccc acctgacgga cctgtaatgc
     2401 aaaagaagac gatgggctgg gaagcgagca cagaacgttt gtatccagaa gacggagtgt
     2461 tgaagggtga catcaaaatg gcgctccgtc ttaaggacgg tggtcgttac ttagcggact
     2521 tcaagactac ctacaaggcc aagaaaccgg tgcaaatgcc cggtgcgtac aacgtggaca
     2581 ggaaactgga cattacatca cataacgaag actataccgt tgtggaacag tacgagagga
     2641 gtgagggccg acactcgaca ggtggcatgg acgagctcta taagttggac ggcgctgggg
     2701 ccggtgctgg ggcaggcgcg atggtgagta aaggtgaaga ggacaacatg gcaagcttgc
     2761 cggcaacaca tgaactgcac atctttgggt caattaatgg tgtagacttc gacatggtgg
     2821 gacagggaac agggaatcct aacgacgggt acgaagaatt aaacctgaaa agcacaaagg
     2881 gagacctaca gttctctccg tggatcctcg tgcctcacat tgggtacggc tttcaccaat
     2941 acctgccgta tccagacggc atgtccccct tccaagcagc gatggtggac ggttccggct
     3001 accaggtcca cagaaccatg cagtttgaag acggtgcatc tcttacagtc aactatcgtt
     3061 acacatatga aggaagtcac attaaagggg aagcacaagt gaaaggtaca gggtttcccg
     3121 cagacggccc agtgatgacg aatagtttaa ctgcggcgga ctggtgccgt tcaaagaaga
     3181 cctatccaaa tgacaagacg attatatcga cgtttaaatg gtcatacact acgggtaatg
     3241 ggaaacgtta taggagcaca gcgcgtacga catatacatt cgcaaaacca atggccgcca
     3301 attaccttaa gaatcaacct atgtacgtgt ttcgaaagac tgagttgaag cactctaaaa
     3361 cggagctcaa tttcaaagag tggcagaagg ccttcacgga cgtgatgggt atggacgagc
     3421 tatataaatc cggactcaga tcacgagctc aagcttcaaa ttctgcagtc gacggtaccg
     3481 cgggcccggg atcatcaaca agtttgtaca aaaaagctga acgagaaacg taaaatgata
     3541 taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa
     3601 cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta cactttatgc
     3661 ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa
     3721 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     3781 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     3841 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     3901 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggagttcc gtatggcaat
     3961 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     4021 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     4081 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     4141 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     4201 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     4261 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     4321 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     4381 cggggcgtaa agatctggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
     4441 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
     4501 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
     4561 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
     4621 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
     4681 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
     4741 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     4801 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     4861 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     4921 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
     4981 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     5041 tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt
     5101 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     5161 atcattttac gtttctcgtt cagctttctt gtacaaagtg gttgatgatc ctaggtgagt
     5221 ctagagagtt aattaagacc cgggactagt ccctagagtc ctgctttaat gagatatgcg
     5281 agacgcctat gatcgcatga tatttgcttt caattctgtt gtgcacgttg taaaaaacct
     5341 gagcatgtgt agctcagatc cttaccgccg gtttcggttc attctaatga atatatcacc
     5401 cgttactatc gtatttttat gaataatatt ctccgttcaa tttactgatt gtaccctact
     5461 acttatatgt acaatattaa aatgaaaaca atatattgtg ctgaataggt ttatagcgac
     5521 atctatgata gagcgccaca ataacaaaca attgcgtttt attattacaa atccaatttt
     5581 aaaaaaagcg gcagaaccgg tcaaacctaa aagactgatt acataaatct tattcaaatt
     5641 tcaaaagtgc cccaggggct agtatctacg acacaccgag cggcgaacta ataacgctca
     5701 ctgaagggaa ctccggttcc ccgccggcgc gcatgggtga gattccttga agttgagtat
     5761 tggccgtccg ctctaccgaa agttacgggc accattcaac ccggtccagc acggcggccg
     5821 ggtaaccgac ttgctgcccc gagaattatg cagcattttt ttggtgtatg tgggccccaa
     5881 atgaagtgca ggtcaaacct tgacagtgac gacaaatcgt tgggcgggtc cagggcgaat
     5941 tttgcgacaa catgtcgagg ctcagcagga cctgcaggtc gactctagag gatccccggg
     6001 taccgagctc gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     6061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8161 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8221 taggcgtatc acgaggccct ttcgtc
//
