LOCUS       sequence-422345-        5705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses NanoLuciferase (Nluc) with the 
            truncated 35sCAMV promoter and Tomato Mosaic Virus UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5705)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 5705)
  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[TMV]:NLuc:NosT
FEATURES             Location/Qualifiers
     source          1..5705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     434..453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             775..1404
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1833..2906
                     /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      2972..3166
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     3629..3649
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        3638..3684
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      4310..4562
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4738..4757)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4749..5564
                     /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             4749..5564
                     /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(4822..4841)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(5650..5705,1..533)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
       61 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      121 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      181 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      241 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      301 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      361 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      421 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      481 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      541 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      601 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga tctctgaaga
      661 aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc
      721 ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag
      781 gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta
      841 gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc
      901 agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac
      961 cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac
     1021 ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg
     1081 attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt
     1141 aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg
     1201 atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc
     1261 attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca
     1321 accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct
     1381 gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa
     1441 acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc
     1501 tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca
     1561 agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca
     1621 tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct
     1681 aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg
     1741 tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg gccctgcaat
     1801 ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca tccggcccgg
     1861 tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg
     1921 cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg
     1981 atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc
     2041 cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc
     2101 gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag
     2161 ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg
     2221 ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg
     2281 aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac
     2341 acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc
     2401 tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg
     2461 ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga
     2521 tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagtt gattggatgt
     2581 accgcgagat cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt
     2641 tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg
     2701 cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca
     2761 agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt
     2821 tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg
     2881 agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag
     2941 caggggaaaa aggtcgaaaa gatctctttc ctgtggatag cacgtacatt gggaacccaa
     3001 agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc
     3061 ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact
     3121 ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc
     3181 gcacagccga agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg
     3241 ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag
     3301 gcaatctgct agcgcgcgga caagctagga ttggcaggat atattcagtt gtaaatgata
     3361 tcgcatgcgg aaacctcctc ggattccatt gcccagctat ctgtcacttt attgagaaga
     3421 tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggccatcg
     3481 ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
     3541 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca
     3601 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
     3661 gaagttcatt tcatttggag aggacacgct cgagtataag agctcatttt tacaacaatt
     3721 accaacaaca acaaacaaca aacaacatta caattacatt tacaattatc gatacacgcg
     3781 tatggtgttt actcttgaag atttcgtggg agactggcgt caaacagcag gctataactt
     3841 agaccaggtc ctagaacagg gaggcgtttc cagtctgttt cagaatcttg gagttagtgt
     3901 tacacctatc caaagaatag ttctcagcgg tgaaaatggc ctcaaaatag acatccatgt
     3961 gatcatccca tacgagggct tgtccggcga tcaaatgggc cagatagaga aaatattcaa
     4021 agtagtgtac ccagtagatg accaccactt taaggtaata cttcactacg ggaccttagt
     4081 tatagacggg gtaaccccta atatgatcga ctacttcggc cgtccctacg agggcattgc
     4141 tgtcttcgat gggaagaaga taacagttac cggcactttg tggaacggta acaaaatcat
     4201 cgacgagcga ctgatcaacc cagatggtag ccttctgttc agggttacca ttaacggggt
     4261 tacagggtgg cgtctgtgcg agcgtatatt agcttaagta tacctgcagg atcgttcaaa
     4321 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     4381 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     4441 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     4501 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     4561 tcggggagct ctggcaggat atatgcggtt gtaattcatt tttattgtct aaatttctgt
     4621 atttgtttgt ttgcaattag atccacgttg tgtctcaaaa tctctgatgt tacattgcac
     4681 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     4741 ggggtgttat gagccatatt caacgggaaa cgtcttgttc gaggccgcga ttaaattcca
     4801 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     4861 cgacaatcta ccgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     4921 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     4981 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     5041 ccactgcgat cccagggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     5101 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     5161 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     5221 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     5281 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     5341 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     5401 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     5461 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     5521 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     5581 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt gttgaatcaa
     5641 ttgatcttct tgaaatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5701 accgc
//
