LOCUS       sequence-422551-        5213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector has an empty backbone with extended multiple 
            cloning site (MSC) where you can inserta a gene into a cassette with
            a strong promoter (truncated 35sCAMV) and UTR (Tomato Mosaic Virus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5213)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 5213)
  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]:MCS+:NosT
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..195
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     658..678
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        667..713
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      847..1099
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1275..1294)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1286..2101
                     /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             1286..2101
                     /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(1359..1378)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2187..2775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2676..2695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3017..3646
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4075..5148
                     /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"
ORIGIN
        1 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
       61 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
      121 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
      181 cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac
      241 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
      301 ccgctcaaaa atggctggcc tacggccagg caatctgcta gcgcgcggac aagctaggat
      361 tggcaggata tattcagttg taaatgatat cgcatgcgga aacctcctcg gattccattg
      421 cccagctatc tgtcacttta ttgagaagat agtggaaaag gaaggtggct cctacaaatg
      481 ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa
      541 agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc
      601 aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta
      661 tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga ggacacgctc
      721 gagtataaga gctcattttt acaacaatta ccaacaacaa caaacaacaa acaacattac
      781 aattacattt acaattatcg atacacgcgt gctcgagggt accgaattcg gatcgtatac
      841 ctgcaggatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
      901 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
      961 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     1021 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     1081 tcatctatgt tactagatcg gggagctctg gcaggatata tgcggttgta attcattttt
     1141 attgtctaaa tttctgtatt tgtttgtttg caattagatc cacgttgtgt ctcaaaatct
     1201 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1261 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgttcgag
     1321 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1381 tgtcgggcaa tcaggtgcga caatctaccg attgtatggg aagcccgatg cgccagagtt
     1441 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     1501 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     1561 tgatgcatgg ttactcacca ctgcgatccc agggaaaaca gcattccagg tattagaaga
     1621 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     1681 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     1741 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     1801 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     1861 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     1921 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     1981 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2041 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2101 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2161 cggctttgtt gaatcaattg atcttcttga aatccttttt ttctgcgcgt aatctgctgc
     2221 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2281 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2341 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2401 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2461 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2521 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2581 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2641 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2701 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2761 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2821 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2881 gcctttgagt gatctctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     2941 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     3001 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     3061 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     3121 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     3181 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     3241 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     3301 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     3361 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     3421 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     3481 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     3541 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     3601 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     3661 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     3721 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     3781 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     3841 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     3901 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     3961 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     4021 ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     4081 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     4141 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     4201 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     4261 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     4321 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     4381 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     4441 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     4501 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     4561 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     4621 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     4681 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     4741 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     4801 atcgagctag ttgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     4861 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     4921 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     4981 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     5041 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     5101 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     5161 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aagatctctt tcc
//
