LOCUS       sequence-443885-        6497 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 Sugarcane bacilliform
            virus (SCBV) promoter and UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6497)
  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/SCBV:nnLuz-I{PIV-216}:NosT
FEATURES             Location/Qualifiers
     source          1..6497
                     /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)"
     intron          3191..3379
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(3634..3645)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      3992..4244
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4420..4439)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4431..5246
                     /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             4431..5246
                     /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(4504..4523)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5332..5920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5821..5840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6162..6497,1..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 atcttttaga agaatatgtc
     2281 caagaacgaa ttgaagtgtt catgattgag aactttgaca agaaaatggg aattcaagat
     2341 gtaccaagta catcaaatgc taacattcca ggaaatttta aatctcttgc agatttgaag
     2401 aaggataaag aagctaaagc tgaatatcaa gacatgcttg ataatcatcg ttcaagcatt
     2461 attgacagac caaggccaac agatgaacac ttcaagcctg gatacatgta caccgattcc
     2521 ctgcagaaga tcaaggagga ctacgcaagc ccaagacagg aggaaccacc atgagaagac
     2581 attgagttct ggttatgcaa ggaagaagac taccacacag aagacctcaa tacagaagat
     2641 gcagttgatc ttactgacgt aagcaatgac gatcagtgga ggcgatcgta agcaatgatg
     2701 cacggaagga caattatgga gcgtggagga cccatcaagc actcagaacg cgaacctcaa
     2761 ctttcggcgc cagcaccttg tatctttagt tggtgtgtgt ctttttcggc atctgtgcca
     2821 ccttaccttt gtcggccacg ttgcctatgc ttagcaccta cgcaagcata gcgctcggct
     2881 ggtgtgtgtt ccctctgcct atataaggca tggttgtaag actcttacac tcatcggtag
     2941 ttcaccacat gatcatttga gcaagtttct cgagatgagg ataaatatct ctttgtctag
     3001 tctctttgag agactgagca aattgtcatc ccgatccatc gcaattacct gcggcgtagt
     3061 cttggcaagt gcaatagcat tcccaattat ccgaagagac tatcaaacgt tccttgaagt
     3121 cggtccgagc tatgccccac agaacttccg aggctatatc atcgtttgtg tcttgtcact
     3181 ttttaggcaa gtaagtttct gcttctacct ttgatatata tataataatt atcattaatt
     3241 agtagtaata taatatttca aatatttttt tcaaaataaa agaatgtagt atatagcaat
     3301 tgcttttctg tagtttataa gtgtgtatat tttaatttat aacttttcta atatatgacc
     3361 aaaatttgtt gatgtgcagg agcagaaagg gttggcaatc tatgacaggc ttccagaaaa
     3421 gaggcgttgg cttgccgact tgccgtttcg tgaagggacg aggccatcca taacctccca
     3481 cattatacag cgtcagcgta ctcagctggt agatcaagag tttgcaacca gagaacttat
     3541 cgataaggtg atcccacgtg tgcaagcaag acacacagac aaaacttttc tcagtacgtc
     3601 aaaatttgag tttcatgcaa aggccatctt ccttctcccc tctatcccta ttaatgatcc
     3661 tcttaacatt ccctcacacg atacggtaag aagaaccaag cgtgagatcg ctcacatgca
     3721 tgattaccat gattgcactc tgcacttggc tcttgctgct caggatggta aggaagtttt
     3781 gaagaagggg tggggccagc gtcacccact ggccggacca ggagtcccag gccctcctac
     3841 tgagtggacc ttcctttacg caccaaggaa cgaagaggag gccagagttg tcgagatgat
     3901 agtcgaagct agtattggct acatgactaa cgatcctgct ggtaagattg tcgaaaatgc
     3961 caagtcaggt tgagaattcg gatccctgca ggatcgttca aacatttggc aataaagttt
     4021 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
     4081 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
     4141 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
     4201 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcggggag ctctggcagg
     4261 atatatgcgg ttgtaattca tttttattgt ctaaatttct gtatttgttt gtttgcaatt
     4321 agatccacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     4381 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     4441 ttcaacggga aacgtcttgt tcgaggccgc gattaaattc caacatggat gctgatttat
     4501 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc taccgattgt
     4561 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     4621 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     4681 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atcccaggga
     4741 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     4801 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     4861 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     4921 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     4981 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     5041 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     5101 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     5161 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     5221 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     5281 gcattacgct gacttgacgg gacggcggct ttgttgaatc aattgatctt cttgaaatcc
     5341 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5401 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5461 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5521 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5581 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5641 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5701 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5761 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5821 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5881 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5941 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6001 tgattctgtg gataaccgta ttaccgcctt tgagtgatct ctgaagaaac cgagcgccgc
     6061 cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat
     6121 atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac
     6181 ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc
     6241 aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt
     6301 gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg
     6361 accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg
     6421 cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc
     6481 caagccctta cgacata
//
