LOCUS       sequence-325398-        7608 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 7608)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7608)
  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            "unpublished"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:MtIDA38:T35s
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    46..70
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        1556..1735
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             1757..2551
                     /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(1811..1830)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2421..2440
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2564..2816
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2911..2935
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             3199..3828
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4257..5330
                     /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      5396..5590
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        5873..5977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5978..6772
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6943..7531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7432..7451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta cccgccaata
       61 tatcctgtca aacactgata gtttaaactc cctcggtcac atgtgcatcc tctagtggag
      121 ggagagagga ttttgaggaa ataattaatt gaattacttg tattattgat aaagtaattt
      181 agataagttg ttagtacaac ttattgcaac tatggataga caaaaatcac ttattttaag
      241 gggagaagta tataacaact tattataaaa tttctgagca gccggcctcc tccaatcaat
      301 catagagagt ggagcccatt ctggaaaacc aaggaacccc caacttgcac tggtgcggtg
      361 gcccaattca aaaaaaacgg ggcccagcaa agtaacgccg ttgtaacgat ttatcaatcc
      421 aaactcaaaa ggcggcgaga tgcgtttaac tccgtgaaat taacaaaccg ccaacaactt
      481 gcaatttgca actaccgttt ccaagaagaa ctcaaccaca caacgtatcc tatcccaaac
      541 cacacgcaac tagtgacgcg tcataaggac acgtgtcaca atttgactgg ttaataattt
      601 caccgctttt gctataaatt acctccaatc cccttagctt cttcacaatt cagttcccaa
      661 ccctaacaat tcttgttcat atcgcttctc tctactttca aggtatgatc caatttctct
      721 cttctttctc tgtaatcctt tcgttgagtt ttgtttccga tcaatcatag gtagttttct
      781 tgtttcgaag catgagatct aggaattttt tgtgattttc caaaattgag atcggtttga
      841 aattgaattt tacagcttga atctcagatc ttgttttatc aatgttttcg atggcttgcg
      901 atgtagatct atgataattg tggttcagtt ttgttaggaa tcgatttcgg tttagcaatt
      961 gcagattaat tagggtttcc aattgaattc ttcagatccg ttatggaatt atgtcaaata
     1021 atttattcaa attggaaatt attgttagat ccactcttaa tctgtttgat ccaagcttca
     1081 attagggttt tcacttgttt caatttcttg ttatggattc tgatttatct gttgatgtta
     1141 gatccactct taatctgttt gatccaagcg ttaattaggg ttttcacttg tttcaatttc
     1201 gtgtgttgga ttctgattta tctgttgttg atgtgattac agaatggctt tctctcataa
     1261 gaagatgctt ttcacagttc aacagatttt gttgatcttg tttaccattt ttattgcttc
     1321 tcctcaatct ggagttgctt gcagaccatt gttagtacat aatgatcaat ggtcgagtga
     1381 atatgggctt gtttggcaat tgcttgcaaa cagccctgcg ccaccgtccg gcggaggaga
     1441 acccactcat ccctgagcta agcttgaatt cttgcggaga gcggagaatt aagggagtca
     1501 cgttatgacc cccgccgatg acgcgggaca agccgtttta cgtttggaac tgacagaacc
     1561 gcaacgttga aggagccact cagccgcggg tttctggagt ttaatgagct aagcacatac
     1621 gtcagaaacc attattgcgc gttcaaaagt cgcctaaggt cactatcagc tagcaaatat
     1681 ttcttgtcaa aaatgctcca ctgacgttcc ataaattccc ctcggtatcc aattagagtc
     1741 tcatattcac tctcaaatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     1801 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     1861 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     1921 tgccctgaat gaactccaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     1981 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2041 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2101 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2161 ccaagcgaaa catcgcatcg agcgaggacg tactcggatg gaagccggtc ttgtcgatca
     2221 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2281 ggcgcggatg cccgacggcg aggatctcgt cgtgacccag ggcgatgcct gcttgccgaa
     2341 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2401 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     2461 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     2521 cttctatcgc cttcttgacg agttcttctg agcttgtcaa gcagatcgtt caaacatttg
     2581 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     2641 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     2701 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     2761 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgacg
     2821 ctggatccga attcggcagg gcccactgca tccaccccag tacattaaaa acgtccgcaa
     2881 tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc
     2941 cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc
     3001 atcagccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
     3061 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
     3121 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
     3181 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
     3241 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
     3301 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
     3361 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
     3421 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
     3481 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
     3541 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
     3601 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
     3661 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
     3721 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
     3781 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
     3841 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
     3901 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
     3961 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
     4021 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
     4081 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
     4141 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
     4201 tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
     4261 gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
     4321 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
     4381 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
     4441 ccggtgcgtc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
     4501 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
     4561 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
     4621 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
     4681 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
     4741 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
     4801 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
     4861 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
     4921 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
     4981 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
     5041 tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc
     5101 tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac
     5161 gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt
     5221 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag
     5281 tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc
     5341 agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaagcttc tttcctgtgg
     5401 atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga
     5461 acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa
     5521 aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg
     5581 cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc
     5641 ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct
     5701 caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc
     5761 cactcgaccg ccggcgccca catcaaggca ccgagctccc atccccccag gggctgcgcc
     5821 cctcggccgc gaacggcctc accccaaaaa tggcagcgct ggccaattcg tgcgcggaac
     5881 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5941 ttgataaatg cttcaataat attgaaaaag gaagagtatg gctaaaatga gaatatcacc
     6001 ggaattgaaa aaactgatcg aaaaataccg ctgcgcaaaa gatacggaag gaatgtctcc
     6061 tgctaaggta tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag
     6121 ccggtataaa gggaccacct atgatgtgga acgggaaaag gacatgatgc tatggctgga
     6181 aggaaagctg cctgttccaa aggtcctgca ctttgaacgg catgatggct ggagcaatct
     6241 gctcatgagt gaggccgatg gcgtcctttg ctcggaagag tatgaagatg aacaaagccc
     6301 tgaaaagatt atcgagctgt atgcggagtg catcaggctc tttcactcca tcgacatatc
     6361 ggattgtccc tatacgaata gcttagacag ccgcttagcc gaattggatt acttactgaa
     6421 taacgatctg gccgatgtgg attgcgaaaa ctgggaagag gacactccat ttaaagatcc
     6481 gcgcgagctg tatgattttt taaagacgga aaagcccgaa gaggaacttg tcttttccca
     6541 cggcgacctg ggagacagca acatctttgt gaaagatggc aaagtaagtg gctttattga
     6601 tcttgggaga agcggcaggg cggacaagtg gtatgacatt gccttctgcg tccggtcgat
     6661 cagggaggat atcggggaag aacagtatgt cgagctattt tttgacttac tggggatcaa
     6721 gcctgattgg gagaaaataa aatattatat tttactggat gaattgtttt agctgtcaga
     6781 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6841 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7021 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7081 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7261 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7501 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7561 cctgtggttg gcacatacaa atggacgaac ggataaacct tttcacgc
//
