LOCUS       sequence-422734-        6095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mNeonGreen containing the PIV intron 
            (216 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-216}:NosT
FEATURES             Location/Qualifiers
     source          1..6095
                     /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"
     intron          1039..1227
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      1729..1981
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2157..2176)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2168..2983
                     /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             2168..2983
                     /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(2241..2260)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3069..3657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3558..3577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3899..4528
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4957..6030
                     /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 atacatcgag ggtaccgaat tcatggtttc caaaggtgag
      841 gaagataata tggctagttt acctgccaca catgagctac acatattcgg cagcataaac
      901 ggtgtagatt ttgatatggt tggacaaggg acaggtaatc ctaatgatgg atatgaggag
      961 ttgaacctca aatctaccaa gggggacctt caattctccc catggatcct agtccctcac
     1021 attggatacg gcttccacgt aagtttctgc ttctaccttt gatatatata taataattat
     1081 cattaattag tagtaatata atatttcaaa tatttttttc aaaataaaag aatgtagtat
     1141 atagcaattg cttttctgta gtttataagt gtgtatattt taatttataa cttttctaat
     1201 atatgaccaa aatttgttga tgtgcagcag tacttaccat acccagatgg aatgtcccct
     1261 ttccaagcag ccatggttga cggttccggg taccaggtac ataggacaat gcaatttgag
     1321 gacggggctt ctctgactgt gaattaccga tatacctatg aaggctctca cataaagggc
     1381 gaagcacaag tgaagggcac cggatttcct gccgacgggc ctgtaatgac taatagcttg
     1441 accgccgcag attggtgcag gtcaaaaaag acctacccaa atgacaagac aataatctct
     1501 acatttaagt ggagctacac tacagggaat ggtaaacgat atagatcaac tgctagaacc
     1561 acatatacct tcgctaaacc tatggcagcc aattatctga agaaccagcc catgtatgtg
     1621 ttccgaaaga ctgaactcaa gcatagcaaa accgaactca acttcaaaga gtggcaaaag
     1681 gctttcactg acgtgatggg aatggacgag ttatataagt aactgcagga tcgttcaaac
     1741 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata
     1801 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt
     1861 atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac
     1921 aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat
     1981 cggggagctc tggcaggata tatgcggttg taattcattt ttattgtcta aatttctgta
     2041 tttgtttgtt tgcaattaga tccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     2101 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2161 gggtgttatg agccatattc aacgggaaac gtcttgttcg aggccgcgat taaattccaa
     2221 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2281 gacaatctac cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2341 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2401 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2461 cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2521 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2581 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2641 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2701 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2761 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2821 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2881 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2941 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3001 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaatcaat
     3061 tgatcttctt gaaatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3121 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3181 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3241 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3301 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3361 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3421 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3481 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3541 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3601 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3661 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3721 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgatctctg
     3781 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc
     3841 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat
     3901 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca
     3961 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca
     4021 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat
     4081 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat
     4141 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt
     4201 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct
     4261 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg
     4321 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct
     4381 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg
     4441 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc
     4501 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc
     4561 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc
     4621 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac
     4681 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa
     4741 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc
     4801 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
     4861 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct gccggccctg
     4921 caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc
     4981 ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag
     5041 gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc
     5101 gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg
     5161 aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc
     5221 acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga
     5281 cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg
     5341 ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta
     5401 accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt
     5461 ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac
     5521 gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag
     5581 aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac
     5641 aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct agttgattgg
     5701 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac
     5761 tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc
     5821 aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag
     5881 ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac
     5941 gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg
     6001 atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc
     6061 ctagcagggg aaaaaggtcg aaaagatctc tttcc
//
