LOCUS       sequence-422956-        6715 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 original 35sCAMV 
            promoter and Tomato mosaic virus UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6715)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6715)
  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/o35s[TMV]:nnLuz-I{PIV-216}:NosT
FEATURES             Location/Qualifiers
     source          1..6715
                     /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        2746..3091
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3034..3054
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     intron          3409..3597
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(3852..3863)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      4210..4462
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4638..4657)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4649..5464
                     /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             4649..5464
                     /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(4722..4741)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5550..6138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6039..6058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6380..6715,1..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 tcgcatgcag attagccttt tcaatttcag
     2281 aaagaatgct aacccacaga tggttagaga ggcttacgca gcaggtctca tcaagacgat
     2341 ctacccgagc aataatctcc aggaaatcaa ataccttccc aagaaggtta aagatgcagt
     2401 caaaagattc aggactaact gcatcaagaa cacagagaaa gatatatttc tcaagatcag
     2461 aagtactatt ccagtatgga cgattcaagg cttgcttcac aaaccaaggc aagtaataga
     2521 gattggagtc tctaaaaagg tagttcccac tgaatcaaag gccatggagt caaagattca
     2581 aatagaggac ctaacagaac tcgccgtaaa gactggcgaa cagttcatac agagtctctt
     2641 acgactcaat gacaagaaga aaatcttcgt caacatggtg gagcacgaca cacttgtcta
     2701 ctccaaaaat atcaaagata cagtctcaga agaccaaagg gcaattgaga cttttcaaca
     2761 aagggtaata tccggaaacc tcctcggatt ccattgccca gctatctgtc actttattgt
     2821 gaagatagtg gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc
     2881 catcgttgaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
     2941 catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat
     3001 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     3061 ataaggaagt tcatttcatt tggagaggac acgctcgagt ataagagctc atttttacaa
     3121 caattaccaa caacaacaaa caacaaacaa cattacaatt acatttacaa ttatcgatac
     3181 atcgagggta ccatgaggat aaatatctct ttgtctagtc tctttgagag actgagcaaa
     3241 ttgtcatccc gatccatcgc aattacctgc ggcgtagtct tggcaagtgc aatagcattc
     3301 ccaattatcc gaagagacta tcaaacgttc cttgaagtcg gtccgagcta tgccccacag
     3361 aacttccgag gctatatcat cgtttgtgtc ttgtcacttt ttaggcaagt aagtttctgc
     3421 ttctaccttt gatatatata taataattat cattaattag tagtaatata atatttcaaa
     3481 tatttttttc aaaataaaag aatgtagtat atagcaattg cttttctgta gtttataagt
     3541 gtgtatattt taatttataa cttttctaat atatgaccaa aatttgttga tgtgcaggag
     3601 cagaaagggt tggcaatcta tgacaggctt ccagaaaaga ggcgttggct tgccgacttg
     3661 ccgtttcgtg aagggacgag gccatccata acctcccaca ttatacagcg tcagcgtact
     3721 cagctggtag atcaagagtt tgcaaccaga gaacttatcg ataaggtgat cccacgtgtg
     3781 caagcaagac acacagacaa aacttttctc agtacgtcaa aatttgagtt tcatgcaaag
     3841 gccatcttcc ttctcccctc tatccctatt aatgatcctc ttaacattcc ctcacacgat
     3901 acggtaagaa gaaccaagcg tgagatcgct cacatgcatg attaccatga ttgcactctg
     3961 cacttggctc ttgctgctca ggatggtaag gaagttttga agaaggggtg gggccagcgt
     4021 cacccactgg ccggaccagg agtcccaggc cctcctactg agtggacctt cctttacgca
     4081 ccaaggaacg aagaggaggc cagagttgtc gagatgatag tcgaagctag tattggctac
     4141 atgactaacg atcctgctgg taagattgtc gaaaatgcca agtcaggttg agaattcgga
     4201 tccctgcagg atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
     4261 ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
     4321 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
     4381 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
     4441 gtgtcatcta tgttactaga tcggggagct ctggcaggat atatgcggtt gtaattcatt
     4501 tttattgtct aaatttctgt atttgtttgt ttgcaattag atccacgttg tgtctcaaaa
     4561 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     4621 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgttc
     4681 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4741 taatgtcggg caatcaggtg cgacaatcta ccgattgtat gggaagcccg atgcgccaga
     4801 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4861 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4921 tgatgatgca tggttactca ccactgcgat cccagggaaa acagcattcc aggtattaga
     4981 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     5041 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     5101 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     5161 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5221 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5281 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5341 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5401 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5461 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     5521 cggcggcttt gttgaatcaa ttgatcttct tgaaatcctt tttttctgcg cgtaatctgc
     5581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6241 accgcctttg agtgatctct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat
     6301 ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac
     6361 aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg
     6421 caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct
     6481 gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca
     6541 accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg
     6601 ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc
     6661 gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg acata
//
