LOCUS       sequence-443882-        7497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mushroom luciferase (nnLuz) containing 
            the PIV intron (216 bp into the gene) with the rice ubiquitin (R. 
            ubi) promoter and UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7497)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 7497)
  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"
COMMENT     Sequence Label: pLSU1/Os-ubi:nnLuz-I{PIV-216}:NosT
FEATURES             Location/Qualifiers
     source          1..7497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             723..1796
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      1862..2056
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2259..3968
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          3007..3968
                     /note="intron upstream of the coding sequence for the rice 
                     polyubiquitin gene RUBQ2"
     intron          4191..4379
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(4634..4645)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      4992..5244
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5420..5439)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5431..6246
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5431..6246
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5504..5523)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6332..6920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6821..6840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(7162..7497,1..294)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
       61 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
      121 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
      181 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
      241 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
      301 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
      361 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
      421 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
      481 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
      541 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
      601 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
      661 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
      721 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
      781 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
      841 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
      901 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
      961 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     1021 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     1081 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     1141 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     1201 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     1261 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     1321 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     1381 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     1441 acatcgagat cgagctagtt gattggatgt accgcgagat cacagaaggc aagaacccgg
     1501 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     1561 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     1621 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     1681 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     1741 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     1801 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa gatctctttc
     1861 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     1921 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     1981 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     2041 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
     2101 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
     2161 gccgctcaaa aatggctggc ctacggccag gcaatctgct agcgcgcgga caagctagga
     2221 ttggcaggat atattcagtt gtaaatgata tcgcatgcat tcgggtcaag gcggaagcca
     2281 gcgcgccacc ccacgtcagc aaatacggag gcgcggggtt gacggcgtca cccggtccta
     2341 acggcgacca acaaaccagc cagaagaaat tacagtaaaa aaaaagtaaa ttgcactttg
     2401 atccaccttt tattacctaa gtctcaattt ggatcaccct taaacctatc ttttcaattt
     2461 gggccgggtt gtggtttgga ctaccatgaa caacttttcg tcatgtctaa cttccctttc
     2521 agcaaacata tgaaccatat atagaggaga tcggccgtat actagagctg atgtgtttaa
     2581 ggtcgttgat tgcacgagaa aaaaaaatcc aaatcgcaac aatagcaaat ttatctggtt
     2641 caaagtgaaa agatatgttt aaaggtagtc caaagtaaaa cttatagata ataaaatgtg
     2701 gtccaaagcg taattcactc aaaaaaaatc aacgagacgt gtaccaaacg gagacaaacg
     2761 gcatcttctc gaaatttccc aaccgctcgc tcgcccgcct cgtcttcccg gaaaccgcgg
     2821 tggtttcagc gtggcggatt ctccaagcag acggagacgt cacggcacgg gactcctccc
     2881 accacccaac cgccataaat accagccccc tcatctcctc tcctcgcatc agctccaccc
     2941 ccgaaaaatt tctccccaat ctcgcgaggc tctcgtcgtc gaatcgaatc ctctcgcgtc
     3001 ctcaaggtac gctgcttctc ctctcctcgc ttcgtttcga ttcgatttcg gacgggtgag
     3061 gttgttttgt tgctagatcc gattggtggt tagggttgtc gatgtgatta tcgtgagatg
     3121 tttaggggtt gtagatctga tggttgtgat ttgggcacgg ttggttcgat aggtggaatc
     3181 gtggttaggt tttgggattg gatgttggtt ctgatgattg gggggaattt ttacggttag
     3241 atgaattgtt ggatgattcg attggggaaa tcggtgtaga tctgttgggg aattgtggaa
     3301 ctagtcatgc ctgagtgatt ggtgcgattt gtagcgtgtt ccatcttgta ggccttgttg
     3361 cgagcatgtt cagatctact gttccgctct tgattgagtt attggtgcca tgggttggtg
     3421 caaacacagg ctttaatatg ttatatctgt tttgtgtttg atgtagatct gtagggtagt
     3481 tcttcttaga catggttcaa ttatgtagct tgtgcgtttc gatttgattt catatgttca
     3541 cagattagat aatgatgaac tcttttaatt aattgtcaat ggtaaatagg aagtcttgtc
     3601 gctatatctg tcataatgat ctcatgttac tatctgccag taatttatgc taagaactat
     3661 attagaatat catgttacaa tctgtagtaa tatcatgtta caatctgtag ttcatctata
     3721 taatctattg tggtaatttc tttttactat ctgtgtgaag attattgcca ctagttcatt
     3781 ctacttattt ctgaagttca ggatacgtgt gctgttacta cctatctgaa tacatgtgtg
     3841 atgtgcctgt tactatcttt ttgaatacat gtatgttctg ttggaatatg tttgctgttt
     3901 gatccgttgt tgtgtcctta atcttgtgct agttcttacc ctatctgttt ggtgattatt
     3961 tcttgcagct cgagatgagg ataaatatct ctttgtctag tctctttgag agactgagca
     4021 aattgtcatc ccgatccatc gcaattacct gcggcgtagt cttggcaagt gcaatagcat
     4081 tcccaattat ccgaagagac tatcaaacgt tccttgaagt cggtccgagc tatgccccac
     4141 agaacttccg aggctatatc atcgtttgtg tcttgtcact ttttaggcaa gtaagtttct
     4201 gcttctacct ttgatatata tataataatt atcattaatt agtagtaata taatatttca
     4261 aatatttttt tcaaaataaa agaatgtagt atatagcaat tgcttttctg tagtttataa
     4321 gtgtgtatat tttaatttat aacttttcta atatatgacc aaaatttgtt gatgtgcagg
     4381 agcagaaagg gttggcaatc tatgacaggc ttccagaaaa gaggcgttgg cttgccgact
     4441 tgccgtttcg tgaagggacg aggccatcca taacctccca cattatacag cgtcagcgta
     4501 ctcagctggt agatcaagag tttgcaacca gagaacttat cgataaggtg atcccacgtg
     4561 tgcaagcaag acacacagac aaaacttttc tcagtacgtc aaaatttgag tttcatgcaa
     4621 aggccatctt ccttctcccc tctatcccta ttaatgatcc tcttaacatt ccctcacacg
     4681 atacggtaag aagaaccaag cgtgagatcg ctcacatgca tgattaccat gattgcactc
     4741 tgcacttggc tcttgctgct caggatggta aggaagtttt gaagaagggg tggggccagc
     4801 gtcacccact ggccggacca ggagtcccag gccctcctac tgagtggacc ttcctttacg
     4861 caccaaggaa cgaagaggag gccagagttg tcgagatgat agtcgaagct agtattggct
     4921 acatgactaa cgatcctgct ggtaagattg tcgaaaatgc caagtcaggt tgagaattcg
     4981 gatccctgca ggatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     5041 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     5101 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     5161 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     5221 cggtgtcatc tatgttacta gatcggggag ctctggcagg atatatgcgg ttgtaattca
     5281 tttttattgt ctaaatttct gtatttgttt gtttgcaatt agatccacgt tgtgtctcaa
     5341 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     5401 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgt
     5461 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     5521 gataatgtcg ggcaatcagg tgcgacaatc taccgattgt atgggaagcc cgatgcgcca
     5581 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     5641 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     5701 cctgatgatg catggttact caccactgcg atcccaggga aaacagcatt ccaggtatta
     5761 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     5821 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     5881 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     5941 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     6001 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     6061 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     6121 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     6181 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     6241 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     6301 gacggcggct ttgttgaatc aattgatctt cttgaaatcc tttttttctg cgcgtaatct
     6361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7021 ttaccgcctt tgagtgatct ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt
     7081 atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa
     7141 acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca
     7201 ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt
     7261 ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat
     7321 caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc
     7381 ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc
     7441 gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacata
//
