LOCUS       sequence-443887-        7336 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 strong Medicago 
            trunculata MtHP promoter and UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 7336)
  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/MtHp:nnLuz-I{PIV-216}:NosT
FEATURES             Location/Qualifiers
     source          1..7336
                     /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          4030..4218
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(4473..4484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      4831..5083
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5259..5278)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5270..6085
                     /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             5270..6085
                     /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(5343..5362)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6171..6759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6660..6679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(7001..7336,1..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 tcgcatgcct cgatcaatag ttcaaaccaa
     2281 aagaaaacaa aaatgaattc aagaattagc caaagcccaa acacatatag ggattaagcc
     2341 accaattatg ataaaaaaaa aaaaagaaga cttttttgca ttcagtcatc gatagaaaat
     2401 gagcttaatg ttatctaaaa attgaagagc tgagcttgtt gtattattaa atatcttgtc
     2461 gtttctttct ttctgtataa cctacgtgca agcaaaccaa atcatcaaca taaatgagcg
     2521 gcgggatttt gaaacacctt ctaaacaact ttttttctaa ataaaactta atgaatgttt
     2581 ccttttttga atctttaaca aaaacactta gaacacttgt taatagtaaa aaatgaagaa
     2641 tataatttgt attttattaa aatttactaa aaatagtatg atcctccttt agattgagtc
     2701 catataagat gtcccgtaag tttaaatata aaaatcatgg tttggctctc tctatcggga
     2761 aaccaaatat gtaaaattga ttttgccata tttgtccgat gcttattgaa attgattttt
     2821 atctctataa ttatataact tgaagttaga acttagaact gcagctttta atttttaata
     2881 tgatttttat actcaaattt agcgttcaag gcacgcggac atgaatatat ttaaatataa
     2941 ctaattcaca tttaacccat ttttaactaa atcattttat ttgaaaaaaa atttgttggc
     3001 aaaactaaac atgcactaaa ccaagttgtt tggcctccgg tggaaaggag ctctttccaa
     3061 ggatgtatcc ggaaaatacc ggactcgatt cacagaggga acaacgcttg accaatttgc
     3121 atgagccaga ttagtcacca acctttggtg ggtcgaaaac tggtgcaaaa gccaaaaaat
     3181 ccattaaaaa aactaaacac gcactaaata tttccaacta aagtattagt attatttaaa
     3241 aaaaaaacaa aaactatatt gtaaaacaaa tagtagtatt atttagtccg gattctagaa
     3301 tggcgtcttg agactgaatt gtttaatatt aaacaaataa aataaaaaat gagagactgc
     3361 aacataattt cctcttcttt tttgggtttg catttgatcg tgtaaaagag aaatctagat
     3421 acaagaatac gcagattttg aatatattgc catagctttt gtggttattt gaatagtata
     3481 tcaggaattt tgaaatttga ctggtcttag aggagataaa agaagaaaaa aaaaatagta
     3541 tatcaagaat tttgtgaacc caaataattt ttttctttaa aattcatatc tcaatgtgaa
     3601 taataaaatg gttccaacaa cctgatatta gttagtcatc atctttatct aattagcaat
     3661 ctaaagtcca aacaatactg taccaaaaaa agtctaaata atacaacttc agttcctata
     3721 aatactagcc ataactacat tcataaacca cacattacga ccattatttg ttatttctta
     3781 cctagcattt taatacatta taccatcctc gagatgagga taaatatctc tttgtctagt
     3841 ctctttgaga gactgagcaa attgtcatcc cgatccatcg caattacctg cggcgtagtc
     3901 ttggcaagtg caatagcatt cccaattatc cgaagagact atcaaacgtt ccttgaagtc
     3961 ggtccgagct atgccccaca gaacttccga ggctatatca tcgtttgtgt cttgtcactt
     4021 tttaggcaag taagtttctg cttctacctt tgatatatat ataataatta tcattaatta
     4081 gtagtaatat aatatttcaa atattttttt caaaataaaa gaatgtagta tatagcaatt
     4141 gcttttctgt agtttataag tgtgtatatt ttaatttata acttttctaa tatatgacca
     4201 aaatttgttg atgtgcagga gcagaaaggg ttggcaatct atgacaggct tccagaaaag
     4261 aggcgttggc ttgccgactt gccgtttcgt gaagggacga ggccatccat aacctcccac
     4321 attatacagc gtcagcgtac tcagctggta gatcaagagt ttgcaaccag agaacttatc
     4381 gataaggtga tcccacgtgt gcaagcaaga cacacagaca aaacttttct cagtacgtca
     4441 aaatttgagt ttcatgcaaa ggccatcttc cttctcccct ctatccctat taatgatcct
     4501 cttaacattc cctcacacga tacggtaaga agaaccaagc gtgagatcgc tcacatgcat
     4561 gattaccatg attgcactct gcacttggct cttgctgctc aggatggtaa ggaagttttg
     4621 aagaaggggt ggggccagcg tcacccactg gccggaccag gagtcccagg ccctcctact
     4681 gagtggacct tcctttacgc accaaggaac gaagaggagg ccagagttgt cgagatgata
     4741 gtcgaagcta gtattggcta catgactaac gatcctgctg gtaagattgt cgaaaatgcc
     4801 aagtcaggtt gagaattcgg atccctgcag gatcgttcaa acatttggca ataaagtttc
     4861 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
     4921 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
     4981 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
     5041 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcggggagc tctggcagga
     5101 tatatgcggt tgtaattcat ttttattgtc taaatttctg tatttgtttg tttgcaatta
     5161 gatccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     5221 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     5281 tcaacgggaa acgtcttgtt cgaggccgcg attaaattcc aacatggatg ctgatttata
     5341 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct accgattgta
     5401 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     5461 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     5521 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tcccagggaa
     5581 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     5641 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     5701 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     5761 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     5821 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     5881 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     5941 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     6001 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     6061 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     6121 cattacgctg acttgacggg acggcggctt tgttgaatca attgatcttc ttgaaatcct
     6181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6301 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6781 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6841 gattctgtgg ataaccgtat taccgccttt gagtgatctc tgaagaaacc gagcgccgcc
     6901 gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata
     6961 tgatgcgatg agtaaataaa caaatacgca aggggaacgc atgaaggtta tcgctgtact
     7021 taaccagaaa ggcgggtcag gcaagacgac catcgcaacc catctagccc gcgccctgca
     7081 actcgccggg gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg
     7141 ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga
     7201 ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc
     7261 ggcggacttg gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc
     7321 aagcccttac gacata
//
