LOCUS       sequence-420442-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning compatible vector for transient expression in 
            protoplasts. Arabidopsis UBQ10 promoter for N-terminal mNeonGreen 
            fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  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 7514)
  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: pLCS107
FEATURES             Location/Qualifiers
     source          1..7514
                     /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    2766..2890
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2915..2920,2921..2938,2939..2945)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2999..3658
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3065..3084)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4000..4305
                     /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     4184..4203
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4346..4470)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4529..5236
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(5293..5309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5293..5309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5306..5328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5317..5333
                     /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(5341..5347,5348..5365,5366..5371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5386..5407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5524..5541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5695..6283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5775..5794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6454..7245,7246..7314))
                     /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     7077..7096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7315..7419)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7487..7505
                     /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 ggttagtaag ggggaagagg ataacatggc atcactcccg gctacccatg
     2041 aactgcacat atttggaagt attaacggcg ttgattttga catggtcggc cagggcacgg
     2101 gaaaccccaa cgacggatat gaggagctta atcttaaaag tacgaaggga gatttgcagt
     2161 tctctccctg gatactggtc ccgcatatcg ggtacggttt ccatcaatat ctgccctacc
     2221 ccgacggtat gagtcctttc caggctgcaa tggtggacgg ctctggctac caagttcata
     2281 ggacgatgca attcgaggac ggtgcaagtc ttacagtcaa ttatagatac acatatgagg
     2341 gttctcacat aaagggagag gcccaggtga aagggacggg cttccccgct gatggccccg
     2401 tgatgacaaa ctcactcacg gcagccgact ggtgtaggtc taaaaagact taccctaacg
     2461 acaagacgat tatatcaaca ttcaaatgga gctacaccac tgggaatggc aagcgttaca
     2521 gaagtactgc caggacaaca tacacttttg cgaagcccat ggctgctaac tatctaaaaa
     2581 accaaccaat gtatgttttc agaaaaaccg agcttaagca cagtaagact gaactaaact
     2641 ttaaagagtg gcaaaaggct tttacggatg tcatgggtat ggacgagctt tacaagtccg
     2701 gactcagatc acgagctcaa gcttcaaatt ctgcagtcga cggtaccgcg ggcccgggat
     2761 catcaacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata
     2821 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     2881 agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2941 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
     3001 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     3061 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     3121 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     3181 cattcttgcc cgcctgatga atgctcatcc ggagttccgt atggcaatga aagacggtga
     3241 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     3301 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     3361 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     3421 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     3481 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     3541 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     3601 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaaag
     3661 atctggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg
     3721 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg
     3781 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg
     3841 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg
     3901 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa
     3961 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc
     4021 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
     4081 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc
     4141 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
     4201 atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
     4261 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt
     4321 atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta
     4381 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     4441 ttctcgttca gctttcttgt acaaagtggt tgatgatcct aggtgagtct agagagttaa
     4501 ttaagacccg ggactagtcc ctagagtcct gctttaatga gatatgcgag acgcctatga
     4561 tcgcatgata tttgctttca attctgttgt gcacgttgta aaaaacctga gcatgtgtag
     4621 ctcagatcct taccgccggt ttcggttcat tctaatgaat atatcacccg ttactatcgt
     4681 atttttatga ataatattct ccgttcaatt tactgattgt accctactac ttatatgtac
     4741 aatattaaaa tgaaaacaat atattgtgct gaataggttt atagcgacat ctatgataga
     4801 gcgccacaat aacaaacaat tgcgttttat tattacaaat ccaattttaa aaaaagcggc
     4861 agaaccggtc aaacctaaaa gactgattac ataaatctta ttcaaatttc aaaagtgccc
     4921 caggggctag tatctacgac acaccgagcg gcgaactaat aacgctcact gaagggaact
     4981 ccggttcccc gccggcgcgc atgggtgaga ttccttgaag ttgagtattg gccgtccgct
     5041 ctaccgaaag ttacgggcac cattcaaccc ggtccagcac ggcggccggg taaccgactt
     5101 gctgccccga gaattatgca gcattttttt ggtgtatgtg ggccccaaat gaagtgcagg
     5161 tcaaaccttg acagtgacga caaatcgttg ggcgggtcca gggcgaattt tgcgacaaca
     5221 tgtcgaggct cagcaggacc tgcaggtcga ctctagagga tccccgggta ccgagctcga
     5281 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5341 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5401 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7441 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     7501 gaggcccttt cgtc
//
