LOCUS       sequence-325081-        8351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of Peptide coding gene (Binary vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8351)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8351)
  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"; journalName: 
            "unpublished"
COMMENT     Sequence Label: LjUBQ:MtNodGRP32:T35s
FEATURES             Location/Qualifiers
     source          1..8351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    328..502
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        602..781
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             803..1597
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(857..876)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1467..1486
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1610..1862
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1957..1981
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2245..2874
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3303..4376
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      4442..4636
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4919..5023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5024..5818
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      5989..6577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6478..6497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6700..6724
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gaggaggaaa catgggaggg ggaggaggaa acatgggagg gagaggagga ggaaacatgg
       61 gagggggagg aggaggagga aatatgggag gttggggagg gggaggaaac atgggaggga
      121 gaggaggagg aaacatggga ggttggggag gaggagaaaa tgaagtagct atgggaagag
      181 gagaatatag tggttatgga ggaggatgtg gcggaggaga atatcaaatg cctcatatga
      241 gaggaggaga acaagaaggc tttgaaaatc aagggccctg ggaatgtcca taagcttctc
      301 tagctagagt cgatcgacaa gctcgagttt ctccataata atgtgtgagt agttcccaga
      361 taagggaatt agggttccta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta
      421 gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa
      481 atccagtact aaaatccaga tcgctaagct tgaattcttg cggagagcgg agaattaagg
      541 gagtcacgtt atgacccccg ccgatgacgc gggacaagcc gttttacgtt tggaactgac
      601 agaaccgcaa cgttgaagga gccactcagc cgcgggtttc tggagtttaa tgagctaagc
      661 acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact atcagctagc
      721 aaatatttct tgtcaaaaat gctccactga cgttccataa attcccctcg gtatccaatt
      781 agagtctcat attcactctc aaatgattga acaagatgga ttgcacgcag gttctccggc
      841 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
      901 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
      961 gtccggtgcc ctgaatgaac tccaagacga ggcagcgcgg ctatcgtggc tggccacgac
     1021 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     1081 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     1141 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     1201 cgaccaccaa gcgaaacatc gcatcgagcg aggacgtact cggatggaag ccggtcttgt
     1261 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     1321 gctcaaggcg cggatgcccg acggcgagga tctcgtcgtg acccagggcg atgcctgctt
     1381 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     1441 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     1501 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     1561 catcgccttc tatcgccttc ttgacgagtt cttctgagct tgtcaagcag atcgttcaaa
     1621 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     1681 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     1741 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     1801 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     1861 tcgacgctgg atccgaattc ggcagggccc actgcatcca ccccagtaca ttaaaaacgt
     1921 ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc
     1981 accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg
     2041 cagcccatca gccgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
     2101 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
     2161 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa
     2221 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
     2281 tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat
     2341 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
     2401 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
     2461 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
     2521 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
     2581 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
     2641 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
     2701 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
     2761 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
     2821 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
     2881 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
     2941 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
     3001 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
     3061 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
     3121 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
     3181 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
     3241 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
     3301 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
     3361 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
     3421 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
     3481 cggcagccgg tgcgtcgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
     3541 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     3601 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     3661 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     3721 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     3781 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     3841 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     3901 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     3961 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     4021 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
     4081 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     4141 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     4201 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     4261 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     4321 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     4381 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa agcttctttc
     4441 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     4501 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     4561 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     4621 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
     4681 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
     4741 gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc
     4801 cgtcgccact cgaccgccgg cgcccacatc aaggcaccga gctcccatcc ccccaggggc
     4861 tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agcgctggcc aattcgtgcg
     4921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4981 ataaccttga taaatgcttc aataatattg aaaaaggaag agtatggcta aaatgagaat
     5041 atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc gcaaaagata cggaaggaat
     5101 gtctcctgct aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac
     5161 ggacagccgg tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg
     5221 gctggaagga aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag
     5281 caatctgctc atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca
     5341 aagccctgaa aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga
     5401 catatcggat tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt
     5461 actgaataac gatctggccg atgtggattg cgaaaactgg gaagaggaca ctccatttaa
     5521 agatccgcgc gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt
     5581 ttcccacggc gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt
     5641 tattgatctt gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg
     5701 gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg
     5761 gatcaagcct gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagct
     5821 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6121 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6601 acggttcctg tggttggcac atacaaatgg acgaacggat aaaccttttc acgccctttt
     6661 aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct
     6721 gtcaaacact gatagtttaa actccctcgg tcacatgtgc atcctctagt ggagggagag
     6781 aggattttga ggaaataatt aattgaatta cttgtattat tgataaagta atttagataa
     6841 gttgttagta caacttattg caactatgga tagacaaaaa tcacttattt taaggggaga
     6901 agtatataac aacttattat aaaatttctg agcagccggc ctcctccaat caatcataga
     6961 gagtggagcc cattctggaa aaccaaggaa cccccaactt gcactggtgc ggtggcccaa
     7021 ttcaaaaaaa acggggccca gcaaagtaac gccgttgtaa cgatttatca atccaaactc
     7081 aaaaggcggc gagatgcgtt taactccgtg aaattaacaa accgccaaca acttgcaatt
     7141 tgcaactacc gtttccaaga agaactcaac cacacaacgt atcctatccc aaaccacacg
     7201 caactagtga cgcgtcataa ggacacgtgt cacaatttga ctggttaata atttcaccgc
     7261 ttttgctata aattacctcc aatcccctta gcttcttcac aattcagttc ccaaccctaa
     7321 caattcttgt tcatatcgct tctctctact ttcaaggtat gatccaattt ctctcttctt
     7381 tctctgtaat cctttcgttg agttttgttt ccgatcaatc ataggtagtt ttcttgtttc
     7441 gaagcatgag atctaggaat tttttgtgat tttccaaaat tgagatcggt ttgaaattga
     7501 attttacagc ttgaatctca gatcttgttt tatcaatgtt ttcgatggct tgcgatgtag
     7561 atctatgata attgtggttc agttttgtta ggaatcgatt tcggtttagc aattgcagat
     7621 taattagggt ttccaattga attcttcaga tccgttatgg aattatgtca aataatttat
     7681 tcaaattgga aattattgtt agatccactc ttaatctgtt tgatccaagc ttcaattagg
     7741 gttttcactt gtttcaattt cttgttatgg attctgattt atctgttgat gttagatcca
     7801 ctcttaatct gtttgatcca agcgttaatt agggttttca cttgtttcaa tttcgtgtgt
     7861 tggattctga tttatctgtt gttgatgtga ttacagaatg aaaatcaaac attttatctt
     7921 cgtgttcttt ctttatgcgc taatccttat ctctgttgtg gcaattaatt cttccaaaga
     7981 tggagaacaa tgtaaatctc tttctatata tatatagaca cacataaatt attttctcaa
     8041 atcctttctt atttaatcat atttaataaa tgtgtttttt tttttatttg cagttggtgc
     8101 aactgaagaa ttcaaaacaa aaattgtggt agatatggat gaaggtttga aaggagttag
     8161 tcatggtgaa gaccatggag gaggacacgt cgatggaggt ttgggatttt ggggaaattt
     8221 gggagggcta aaaggttggt ttgaatggat ggcaaacgaa gcaatttcgg gaataggaga
     8281 aagccaagga gggtttggag tcagttgggg aggaggagga aacatgggag ggggaggagg
     8341 aaacatggga g
//
