LOCUS       sequence-422962-        6102 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 truncated 35sCAMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6102)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6102)
  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/t35s[]:nnLuz-I{PIV-216}:NosT
FEATURES             Location/Qualifiers
     source          1..6102
                     /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)"
     primer_bind     2519..2539
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        2528..2573
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     intron          2796..2984
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(3239..3250)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      3597..3849
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4025..4044)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4036..4851
                     /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             4036..4851
                     /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(4109..4128)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4937..5525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5426..5445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5767..6102,1..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 tcgcatgcgg aaacctcctc ggattccatt
     2281 gcccagctat ctgtcacttt attgagaaga tagtggaaaa ggaaggtggc tcctacaaat
     2341 gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca
     2401 aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt
     2461 caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact
     2521 atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag aggctcgaga
     2581 tgaggataaa tatctctttg tctagtctct ttgagagact gagcaaattg tcatcccgat
     2641 ccatcgcaat tacctgcggc gtagtcttgg caagtgcaat agcattccca attatccgaa
     2701 gagactatca aacgttcctt gaagtcggtc cgagctatgc cccacagaac ttccgaggct
     2761 atatcatcgt ttgtgtcttg tcacttttta ggcaagtaag tttctgcttc tacctttgat
     2821 atatatataa taattatcat taattagtag taatataata tttcaaatat ttttttcaaa
     2881 ataaaagaat gtagtatata gcaattgctt ttctgtagtt tataagtgtg tatattttaa
     2941 tttataactt ttctaatata tgaccaaaat ttgttgatgt gcaggagcag aaagggttgg
     3001 caatctatga caggcttcca gaaaagaggc gttggcttgc cgacttgccg tttcgtgaag
     3061 ggacgaggcc atccataacc tcccacatta tacagcgtca gcgtactcag ctggtagatc
     3121 aagagtttgc aaccagagaa cttatcgata aggtgatccc acgtgtgcaa gcaagacaca
     3181 cagacaaaac ttttctcagt acgtcaaaat ttgagtttca tgcaaaggcc atcttccttc
     3241 tcccctctat ccctattaat gatcctctta acattccctc acacgatacg gtaagaagaa
     3301 ccaagcgtga gatcgctcac atgcatgatt accatgattg cactctgcac ttggctcttg
     3361 ctgctcagga tggtaaggaa gttttgaaga aggggtgggg ccagcgtcac ccactggccg
     3421 gaccaggagt cccaggccct cctactgagt ggaccttcct ttacgcacca aggaacgaag
     3481 aggaggccag agttgtcgag atgatagtcg aagctagtat tggctacatg actaacgatc
     3541 ctgctggtaa gattgtcgaa aatgccaagt caggttgaga attcggatcc ctgcaggatc
     3601 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     3661 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     3721 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     3781 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     3841 tactagatcg gggagctctg gcaggatata tgcggttgta attcattttt attgtctaaa
     3901 tttctgtatt tgtttgtttg caattagatc cacgttgtgt ctcaaaatct ctgatgttac
     3961 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     4021 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgttcgag gccgcgatta
     4081 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     4141 tcaggtgcga caatctaccg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     4201 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     4261 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     4321 ttactcacca ctgcgatccc agggaaaaca gcattccagg tattagaaga atatcctgat
     4381 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     4441 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     4501 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     4561 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     4621 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     4681 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     4741 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     4801 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4861 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt
     4921 gaatcaattg atcttcttga aatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5161 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5221 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5281 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5341 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5401 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5461 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5521 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5581 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5641 gatctctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag
     5701 cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg
     5761 gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc
     5821 gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc
     5881 gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt
     5941 gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc
     6001 gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc
     6061 gacttcgtgc tgattccggt gcagccaagc ccttacgaca ta
//
