LOCUS       sequence-422736-        6095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mNeonGreen containing the PIV intron 
            (390 bp into the gene) with extra 3' splice site removed (corrected 
            intron) with the truncated 35sCAMV promoter and TMV UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6095)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6095)
  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]:mNeonGreen-CI{PIV-390}:NosT
FEATURES             Location/Qualifiers
     source          1..6095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     506..526
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        515..561
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     intron          1061..1249
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      1577..1829
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2005..2024)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2016..2831
                     /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             2016..2831
                     /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(2089..2108)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2917..3505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3406..3425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3747..4376
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4805..5878
                     /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      join(5944..6095,1..43)
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca
       61 cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg
      121 cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca
      181 atctgctagc gcgcggacaa gctaggattg gcaggatata ttcagttgta aatgatatcg
      241 catgcggaaa cctcctcgga ttccattgcc cagctatctg tcactttatt gagaagatag
      301 tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag gccatcgttg
      361 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
      421 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg
      481 acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa
      541 gttcatttca tttggagagg acacgctcga gtataagagc tcatttttac aacaattacc
      601 aacaacaaca aacaacaaac aacattacaa ttacatttac aattatcgat acatcgaggg
      661 taccgaattc atggtttcca aaggtgagga agataatatg gctagtttac ctgccacaca
      721 tgagctacac atattcggca gcataaacgg tgtagatttt gatatggttg gacaagggac
      781 aggtaatcct aatgatggat atgaggagtt gaacctcaaa tctaccaagg gggaccttca
      841 attctcccca tggatcctag tccctcacat tggatacggc ttccaccagt acttaccata
      901 cccagatgga atgtcccctt tccaagcagc catggttgac ggttccgggt accaggtaca
      961 taggacaatg caatttgagg acggggcttc tctgactgtg aattaccgat atacctatga
     1021 aggctctcac ataaagggcg aagcacaagt gaagggcacc gtaagtttct gcttctacct
     1081 ttgatatata tataataatt atcattaatt agtagtaata taatatttca aatatttttt
     1141 tcaaaataaa agaatgtagt atatagcaat tgcttttctg tagtttataa gtgtgtatat
     1201 tttaatttat aacttttcta atatatgacc aaaatttgtt gatgtgcagg gatttcctgc
     1261 cgacgggcct gtaatgacta atagcttgac cgccgcagat tggtgcaggt caaaaaagac
     1321 ctacccaaat gacaagacaa taatctctac atttaagtgg agctacacta cagggaatgg
     1381 taaacgatat agatcaactg ctagaaccac atataccttc gctaaaccta tggcagccaa
     1441 ttatctgaag aaccagccca tgtatgtgtt ccgaaagact gaactcaagc atagcaaaac
     1501 cgaactcaac ttcaaagagt ggcaaaaggc tttcactgac gtgatgggaa tggacgagtt
     1561 atataagtaa ctgcaggatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     1621 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     1681 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     1741 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     1801 gcgcgcggtg tcatctatgt tactagatcg gggagctctg gcaggatata tgcggttgta
     1861 attcattttt attgtctaaa tttctgtatt tgtttgtttg caattagatc cacgttgtgt
     1921 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1981 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     2041 cttgttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2101 ctcgcgataa tgtcgggcaa tcaggtgcga caatctaccg attgtatggg aagcccgatg
     2161 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2221 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2281 gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca gcattccagg
     2341 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     2401 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2461 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2521 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     2581 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2641 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2701 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2761 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2821 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     2881 gacgggacgg cggctttgtt gaatcaattg atcttcttga aatccttttt ttctgcgcgt
     2941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3061 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3541 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3601 ccgtattacc gcctttgagt gatctctgaa gaaaccgagc gccgccgtct aaaaaggtga
     3661 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
     3721 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
     3781 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
     3841 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
     3901 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
     3961 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
     4021 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
     4081 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
     4141 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
     4201 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
     4261 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
     4321 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
     4381 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
     4441 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
     4501 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
     4561 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
     4621 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
     4681 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
     4741 taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat
     4801 cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga
     4861 cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc
     4921 acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc
     4981 gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt
     5041 tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc
     5101 cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc
     5161 agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga
     5221 cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa
     5281 gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg
     5341 gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac
     5401 cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc
     5461 cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga
     5521 gtacatcgag atcgagctag ttgattggat gtaccgcgag atcacagaag gcaagaaccc
     5581 ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct
     5641 ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat
     5701 ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct
     5761 gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc
     5821 gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg
     5881 tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aagatctctt
     5941 tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta
     6001 cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa
     6061 agagaaaaaa ggcgattttt ccgcctaaaa ctctt
//
