LOCUS       sequence-422717-        6003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mNeonGreen containing the PIV intron 
            (198bp into the gene) with extra 3' splice site removed (corrected 
            intron) with the truncated 35sCAMV promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6003)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6003)
  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[]:mNeonGreen-CI{PIV-198}:NosT
FEATURES             Location/Qualifiers
     source          1..6003
                     /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..712
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     intron          917..1105
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      1637..1889
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2065..2084)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2076..2891
                     /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             2076..2891
                     /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(2149..2168)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2977..3565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3466..3485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3807..4436
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4865..5938
                     /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 ggctcgagat
      721 ggtttccaaa ggtgaggaag ataatatggc tagtttacct gccacacatg agctacacat
      781 attcggcagc ataaacggtg tagattttga tatggttgga caagggacag gtaatcctaa
      841 tgatggatat gaggagttga acctcaaatc taccaagggg gaccttcaat tctccccatg
      901 gatcctagtc cctcacgtaa gtttctgctt ctacctttga tatatatata ataattatca
      961 ttaattagta gtaatataat atttcaaata tttttttcaa aataaaagaa tgtagtatat
     1021 agcaattgct tttctgtagt ttataagtgt gtatatttta atttataact tttctaatat
     1081 atgaccaaaa tttgttgatg tgcagattgg atacggcttc caccagtact taccataccc
     1141 agatggaatg tcccctttcc aagcagccat ggttgacggt tccgggtacc aggtacatag
     1201 gacaatgcaa tttgaggacg gggcttctct gactgtgaat taccgatata cctatgaagg
     1261 ctctcacata aagggcgaag cacaagtgaa gggcaccgga tttcctgccg acgggcctgt
     1321 aatgactaat agcttgaccg ccgcagattg gtgcaggtca aaaaagacct acccaaatga
     1381 caagacaata atctctacat ttaagtggag ctacactaca gggaatggta aacgatatag
     1441 atcaactgct agaaccacat ataccttcgc taaacctatg gcagccaatt atctgaagaa
     1501 ccagcccatg tatgtgttcc gaaagactga actcaagcat agcaaaaccg aactcaactt
     1561 caaagagtgg caaaaggctt tcactgacgt gatgggaatg gacgagttat ataagtaaga
     1621 attcggatcc ctgcaggatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     1681 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     1741 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     1801 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     1861 gcgcgcggtg tcatctatgt tactagatcg gggagctctg gcaggatata tgcggttgta
     1921 attcattttt attgtctaaa tttctgtatt tgtttgtttg caattagatc cacgttgtgt
     1981 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     2041 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     2101 cttgttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2161 ctcgcgataa tgtcgggcaa tcaggtgcga caatctaccg attgtatggg aagcccgatg
     2221 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2281 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2341 gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca gcattccagg
     2401 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     2461 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2521 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2581 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     2641 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2701 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2761 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2821 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2881 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     2941 gacgggacgg cggctttgtt gaatcaattg atcttcttga aatccttttt ttctgcgcgt
     3001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3121 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3601 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3661 ccgtattacc gcctttgagt gatctctgaa gaaaccgagc gccgccgtct aaaaaggtga
     3721 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
     3781 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
     3841 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
     3901 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
     3961 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
     4021 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
     4081 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
     4141 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
     4201 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
     4261 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
     4321 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
     4381 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
     4441 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
     4501 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
     4561 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
     4621 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
     4681 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
     4741 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
     4801 taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat
     4861 cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga
     4921 cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc
     4981 acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc
     5041 gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt
     5101 tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc
     5161 cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc
     5221 agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga
     5281 cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa
     5341 gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg
     5401 gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac
     5461 cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc
     5521 cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga
     5581 gtacatcgag atcgagctag ttgattggat gtaccgcgag atcacagaag gcaagaaccc
     5641 ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct
     5701 ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat
     5761 ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct
     5821 gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc
     5881 gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg
     5941 tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aagatctctt
     6001 tcc
//
