LOCUS       sequence-422825-        6523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mNeonGreen containing the PIV intron 
            (198bp into the gene) with the truncated 35sCAMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6523)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6523)
  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/o35s[]:mNeonGreen-I{PIV-198}:NosT
FEATURES             Location/Qualifiers
     source          1..6523
                     /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)"
     promoter        885..1230
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1173..1193
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     intron          1437..1625
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      2157..2409
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2585..2604)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2596..3411
                     /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             2596..3411
                     /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(2669..2688)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3497..4085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3986..4005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4327..4956
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5385..6458
                     /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 cgcatgcaga ttagcctttt caatttcaga
      421 aagaatgcta acccacagat ggttagagag gcttacgcag caggtctcat caagacgatc
      481 tacccgagca ataatctcca ggaaatcaaa taccttccca agaaggttaa agatgcagtc
      541 aaaagattca ggactaactg catcaagaac acagagaaag atatatttct caagatcaga
      601 agtactattc cagtatggac gattcaaggc ttgcttcaca aaccaaggca agtaatagag
      661 attggagtct ctaaaaaggt agttcccact gaatcaaagg ccatggagtc aaagattcaa
      721 atagaggacc taacagaact cgccgtaaag actggcgaac agttcataca gagtctctta
      781 cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac acttgtctac
      841 tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac ttttcaacaa
      901 agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca ctttattgtg
      961 aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggcc
     1021 atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
     1081 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc
     1141 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
     1201 taaggaagtt catttcattt ggagagaaca cgctcgagat ggtttccaaa ggtgaggaag
     1261 ataatatggc tagtttacct gccacacatg agctacacat attcggcagc ataaacggtg
     1321 tagattttga tatggttgga caagggacag gtaatcctaa tgatggatat gaggagttga
     1381 acctcaaatc taccaagggg gaccttcaat tctccccatg gatcctagtc cctcacgtaa
     1441 gtttctgctt ctacctttga tatatatata ataattatca ttaattagta gtaatataat
     1501 atttcaaata tttttttcaa aataaaagaa tgtagtatat agcaattgct tttctgtagt
     1561 ttataagtgt gtatatttta atttataact tttctaatat atgaccaaaa tttgttgatg
     1621 tgcagattgg atacggcttc caccagtact taccataccc agatggaatg tcccctttcc
     1681 aagcagccat ggttgacggt tccgggtacc aggtacatag gacaatgcaa tttgaggacg
     1741 gggcttctct gactgtgaat taccgatata cctatgaagg cagtcacata aagggcgaag
     1801 cacaagtgaa gggcaccgga tttcctgccg acgggcctgt aatgactaat agcttgaccg
     1861 ccgcagattg gtgcaggtca aaaaagacct acccaaatga caagacaata atctctacat
     1921 ttaagtggag ctacactaca gggaatggta aacgatatag atcaactgct agaaccacat
     1981 ataccttcgc taaacctatg gcagccaatt atctgaagaa ccagcccatg tatgtgttcc
     2041 gaaagactga actcaagcat agcaaaaccg aactcaactt caaagagtgg caaaaggctt
     2101 tcactgacgt gatgggaatg gacgagttat ataagtaaga attcggatcc ctgcaggatc
     2161 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     2221 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     2281 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     2341 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     2401 tactagatcg gggagctctg gcaggatata tgcggttgta attcattttt attgtctaaa
     2461 tttctgtatt tgtttgtttg caattagatc cacgttgtgt ctcaaaatct ctgatgttac
     2521 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2581 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgttcgag gccgcgatta
     2641 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2701 tcaggtgcga caatctaccg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2761 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2821 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2881 ttactcacca ctgcgatccc agggaaaaca gcattccagg tattagaaga atatcctgat
     2941 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3001 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     3061 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3121 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3181 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3241 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3301 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3361 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     3421 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt
     3481 gaatcaattg atcttcttga aatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3541 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3601 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3661 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3721 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3781 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3841 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3901 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3961 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4021 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4081 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4141 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4201 gatctctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag
     4261 cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg
     4321 gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc
     4381 gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc
     4441 gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt
     4501 gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc
     4561 gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc
     4621 gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg
     4681 gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc
     4741 gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg
     4801 tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc
     4861 gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag
     4921 gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga
     4981 gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa
     5041 cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg
     5101 aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc
     5161 tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga
     5221 attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg
     5281 gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgtctgc
     5341 cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac
     5401 catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg
     5461 ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc
     5521 aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt
     5581 cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg
     5641 acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc
     5701 gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt
     5761 ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt
     5821 cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg
     5881 tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc
     5941 agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc
     6001 gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta
     6061 gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag
     6121 ttgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc
     6181 ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg
     6241 ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg
     6301 ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc
     6361 cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc
     6421 gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc
     6481 aaattgccct agcaggggaa aaaggtcgaa aagatctctt tcc
//
