LOCUS       sequence-443888-        5964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mCherry containing the PIV intron 
            (180bp into the gene) with the truncated 35sCAMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5964)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 5964)
  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[]:mCherry-I{PIV-180}:NosT
FEATURES             Location/Qualifiers
     source          1..5964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             723..1796
                     /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      1862..2056
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     2519..2539
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        2528..2573
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     intron          2763..2951
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      3459..3711
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3887..3906)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3898..4713
                     /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             3898..4713
                     /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(3971..3990)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4799..5387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5288..5307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5629..5964,1..294)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
       61 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
      121 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
      181 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
      241 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
      301 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
      361 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
      421 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
      481 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
      541 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
      601 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
      661 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
      721 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
      781 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
      841 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
      901 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
      961 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     1021 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     1081 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     1141 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     1201 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     1261 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     1321 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     1381 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     1441 acatcgagat cgagctagtt gattggatgt accgcgagat cacagaaggc aagaacccgg
     1501 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     1561 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     1621 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     1681 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     1741 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     1801 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa gatctctttc
     1861 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     1921 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     1981 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     2041 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
     2101 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
     2161 gccgctcaaa aatggctggc ctacggccag gcaatctgct agcgcgcgga caagctagga
     2221 ttggcaggat atattcagtt gtaaatgata tcgcatgcgg aaacctcctc ggattccatt
     2281 gcccagctat ctgtcacttt attgagaaga tagtggaaaa ggaaggtggc tcctacaaat
     2341 gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca
     2401 aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt
     2461 caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact
     2521 atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag aggcctacgc
     2581 gtatggccat catcaaggag ttcatgcggt tcaaggtgca catggaaggc agcgtgaacg
     2641 gccacgagtt cgagatcgag ggcgagggcg aaggcaggcc ctacgagggc acccagaccg
     2701 ccaagctgaa ggtgaccaag ggcggacctc tgcccttcgc ctgggacatc ctgagccccc
     2761 aggtaagttt ctgcttctac ctttgatata tatataataa ttatcattaa ttagtagtaa
     2821 tataatattt caaatatttt tttcaaaata aaagaatgta gtatatagca attgcttttc
     2881 tgtagtttat aagtgtgtat attttaattt ataacttttc taatatatga ccaaaatttg
     2941 ttgatgtgca gttcatgtac ggcagcaagg cctacgtgaa gcaccccgcc gacatccccg
     3001 actacctgaa gctgtccttc cccgagggat tcaaatggga gcgggtgatg aacttcgagg
     3061 acggcggcgt ggtgaccgtg acccaggaca gcagcctgca ggacggcgag ttcatctaca
     3121 aggtgaaact gcggggcacc aacttcccca gcgacggccc cgtgatgcag aaaaagacca
     3181 tgggctggga ggccagcagc gagcggatgt accccgagga tggcgccctg aagggcgaga
     3241 tcaagcagcg gctgaagctg aaggatggcg gccactacga cgccgaggtg aaaaccacct
     3301 acaaggccaa gaaacccgtg cagctgcctg gcgcctacaa cgtgaacatc aagctggaca
     3361 tcaccagcca caacgaggac tacaccatcg tggagcagta cgagcgggcc gagggcagac
     3421 acagcaccgg cggcatggac gagtaagtat acctgcagga tcgttcaaac atttggcaat
     3481 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     3541 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     3601 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     3661 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cggggagctc
     3721 tggcaggata tatgcggttg taattcattt ttattgtcta aatttctgta tttgtttgtt
     3781 tgcaattaga tccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     3841 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     3901 agccatattc aacgggaaac gtcttgttcg aggccgcgat taaattccaa catggatgct
     3961 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctac
     4021 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     4081 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     4141 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     4201 ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     4261 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     4321 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     4381 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     4441 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     4501 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     4561 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     4621 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     4681 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     4741 tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaatcaat tgatcttctt
     4801 gaaatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5401 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5461 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgatctctg aagaaaccga
     5521 gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc
     5581 tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc
     5641 gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc
     5701 gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc
     5761 cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg
     5821 acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg
     5881 ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg
     5941 gtgcagccaa gcccttacga cata
//
