LOCUS       sequence-325461-        8195 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 8195)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8195)
  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            "unpublished"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:MtLegin6:T35s
FEATURES             Location/Qualifiers
     source          1..8195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    130..154
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    1953..2127
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        2227..2406
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2428..3222
                     /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(2482..2501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3092..3111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3235..3487
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3582..3606
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             3870..4499
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4928..6001
                     /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      6067..6261
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        6544..6648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6649..7443
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      join(7614..8195,1..7)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8103..8122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atggaaaaac gccagcaacg cggccttttt acggttcctg tggttggcac atacaaatgg
       61 acgaacggat aaaccttttc acgccctttt aaatatccga ttattctaat aaacgctctt
      121 ttctcttagg tttacccgcc aatatatcct gtcaaacact gatagtttaa actccctcgg
      181 tcacatgtgc atcctctagt ggagggagag aggattttga ggaaataatt aattgaatta
      241 cttgtattat tgataaagta atttagataa gttgttagta caacttattg caactatgga
      301 tagacaaaaa tcacttattt taaggggaga agtatataac aacttattat aaaatttctg
      361 agcagccggc ctcctccaat caatcataga gagtggagcc cattctggaa aaccaaggaa
      421 cccccaactt gcactggtgc ggtggcccaa ttcaaaaaaa acggggccca gcaaagtaac
      481 gccgttgtaa cgatttatca atccaaactc aaaaggcggc gagatgcgtt taactccgtg
      541 aaattaacaa accgccaaca acttgcaatt tgcaactacc gtttccaaga agaactcaac
      601 cacacaacgt atcctatccc aaaccacacg caactagtga cgcgtcataa ggacacgtgt
      661 cacaatttga ctggttaata atttcaccgc ttttgctata aattacctcc aatcccctta
      721 gcttcttcac aattcagttc ccaaccctaa caattcttgt tcatatcgct tctctctact
      781 ttcaaggtat gatccaattt ctctcttctt tctctgtaat cctttcgttg agttttgttt
      841 ccgatcaatc ataggtagtt ttcttgtttc gaagcatgag atctaggaat tttttgtgat
      901 tttccaaaat tgagatcggt ttgaaattga attttacagc ttgaatctca gatcttgttt
      961 tatcaatgtt ttcgatggct tgcgatgtag atctatgata attgtggttc agttttgtta
     1021 ggaatcgatt tcggtttagc aattgcagat taattagggt ttccaattga attcttcaga
     1081 tccgttatgg aattatgtca aataatttat tcaaattgga aattattgtt agatccactc
     1141 ttaatctgtt tgatccaagc ttcaattagg gttttcactt gtttcaattt cttgttatgg
     1201 attctgattt atctgttgat gttagatcca ctcttaatct gtttgatcca agcgttaatt
     1261 agggttttca cttgtttcaa tttcgtgtgt tggattctga tttatctgtt gttgatgtga
     1321 ttacagaatg acttatgtta agctagctcc ttttgctgtc ttcttgcttg ccgcattctg
     1381 tatgtatttc aatcatcttc ctcacccata tataacatgc atgcatttct atttatgttt
     1441 aataaatcgt gagaaattaa tcataatcga ctctcaacta attttattac tattataata
     1501 atggtatttg tatgttgttc aaatttaatt agctttgtca tttacagtaa tgtttccgat
     1561 gaacaaagta gaaggagcag gatgtcaaca aagttgcggt acctttttgc caggttgtcg
     1621 taacggttgt gaatgcgtac tcgttgattg tacgttcggt gtttgtgtgg gtttttgcga
     1681 aaaagacaat gtggtaagtg gaaattttca caggaaggtt gacgaacatc ctaagttatg
     1741 tatgtctcat gatgaatgca cgaaaaaggg tagtgggaat tattgtgctc actttccaaa
     1801 ttctaatctc aaacatggct tctgttttgc ctctgtctct gaagcataag acgcatacaa
     1861 gatggcctct agtaataaat tcaagaatga cttcttgaag atgtctcttc ctgcttaagc
     1921 ttctctagct agagtcgatc gacaagctcg agtttctcca taataatgtg tgagtagttc
     1981 ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca tataagaaac
     2041 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
     2101 ccaaaatcca gtactaaaat ccagatcgct aagcttgaat tcttgcggag agcggagaat
     2161 taagggagtc acgttatgac ccccgccgat gacgcgggac aagccgtttt acgtttggaa
     2221 ctgacagaac cgcaacgttg aaggagccac tcagccgcgg gtttctggag tttaatgagc
     2281 taagcacata cgtcagaaac cattattgcg cgttcaaaag tcgcctaagg tcactatcag
     2341 ctagcaaata tttcttgtca aaaatgctcc actgacgttc cataaattcc cctcggtatc
     2401 caattagagt ctcatattca ctctcaaatg attgaacaag atggattgca cgcaggttct
     2461 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     2521 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     2581 gacctgtccg gtgccctgaa tgaactccaa gacgaggcag cgcggctatc gtggctggcc
     2641 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     2701 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     2761 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     2821 ccattcgacc accaagcgaa acatcgcatc gagcgaggac gtactcggat ggaagccggt
     2881 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     2941 gccaggctca aggcgcggat gcccgacggc gaggatctcg tcgtgaccca gggcgatgcc
     3001 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3061 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3121 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3181 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcttgtca agcagatcgt
     3241 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     3301 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     3361 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     3421 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     3481 ctagatcgac gctggatccg aattcggcag ggcccactgc atccacccca gtacattaaa
     3541 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
     3601 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
     3661 acaggcagcc catcagccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
     3721 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
     3781 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
     3841 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
     3901 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
     3961 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
     4021 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
     4081 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
     4141 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg
     4201 acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg
     4261 gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg
     4321 aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc
     4381 gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg
     4441 gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag
     4501 ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag
     4561 cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg
     4621 ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca
     4681 aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag
     4741 caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag
     4801 aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag
     4861 gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc aagcccgagg
     4921 aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga
     4981 tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga
     5041 agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca
     5101 accgccggca gccggtgcgt cgtcgattag gaagccgccc aagggcgacg agcaaccaga
     5161 ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt
     5221 ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct
     5281 tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta
     5341 cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg
     5401 gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg
     5461 ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa
     5521 caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt
     5581 atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc
     5641 ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa
     5701 cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt
     5761 tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac
     5821 gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa
     5881 gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg
     5941 cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta
     6001 atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaagctt
     6061 ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc
     6121 gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat
     6181 aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa
     6241 aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc
     6301 gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc
     6361 cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc
     6421 cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accgagctcc catcccccca
     6481 ggggctgcgc ccctcggccg cgaacggcct caccccaaaa atggcagcgc tggccaattc
     6541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6601 agacaataac cttgataaat gcttcaataa tattgaaaaa ggaagagtat ggctaaaatg
     6661 agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgcaaa agatacggaa
     6721 ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct atatttaaaa
     6781 atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa ggacatgatg
     6841 ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg gcatgatggc
     6901 tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga gtatgaagat
     6961 gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct ctttcactcc
     7021 atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc cgaattggat
     7081 tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga ggacactcca
     7141 tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga agaggaactt
     7201 gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg caaagtaagt
     7261 ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat tgccttctgc
     7321 gtccggtcga tcagggagga tatcggggaa gaacagtatg tcgagctatt ttttgactta
     7381 ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga tgaattgttt
     7441 tagctgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7501 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7561 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7621 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7681 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7741 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     7801 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7861 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7921 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7981 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8041 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8101 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8161 cgatttttgt gatgctcgtc aggggggcgg agcct
//
