LOCUS       sequence-325322-        8483 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 8483)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8483)
  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            "unpublished"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:MtnsLTP55:T35s
FEATURES             Location/Qualifiers
     source          1..8483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        140..244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             245..1039
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1210..1798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1699..1718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1921..1945
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    4032..4206
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        4306..4485
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4507..5301
                     /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(4561..4580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5171..5190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5314..5566
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5661..5685
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5949..6578
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7007..8080
                     /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      8146..8340
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccg
       61 agctcccatc cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg
      121 cagcgctggc caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      181 tatgtatccg ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa
      241 gagtatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg
      301 cgcaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga
      361 aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg
      421 ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt
      481 tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc
      541 ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat
      601 caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg
      661 cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg
      721 ggaagaggac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa
      781 gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa
      841 agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta
      901 tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga
      961 gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt
     1021 actggatgaa ttgttttagc tgtcagacca agtttactca tatatacttt agattgattt
     1081 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1321 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1441 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1501 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1561 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1621 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1681 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1741 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1801 cgccagcaac gcggcctttt tacggttcct gtggttggca catacaaatg gacgaacgga
     1861 taaacctttt cacgcccttt taaatatccg attattctaa taaacgctct tttctcttag
     1921 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactccctcg gtcacatgtg
     1981 catcctctag tggagggaga gaggattttg aggaaataat taattgaatt acttgtatta
     2041 ttgataaagt aatttagata agttgttagt acaacttatt gcaactatgg atagacaaaa
     2101 atcacttatt ttaaggggag aagtatataa caacttatta taaaatttct gagcagccgg
     2161 cctcctccaa tcaatcatag agagtggagc ccattctgga aaaccaagga acccccaact
     2221 tgcactggtg cggtggccca attcaaaaaa aacggggccc agcaaagtaa cgccgttgta
     2281 acgatttatc aatccaaact caaaaggcgg cgagatgcgt ttaactccgt gaaattaaca
     2341 aaccgccaac aacttgcaat ttgcaactac cgtttccaag aagaactcaa ccacacaacg
     2401 tatcctatcc caaaccacac gcaactagtg acgcgtcata aggacacgtg tcacaatttg
     2461 actggttaat aatttcaccg cttttgctat aaattacctc caatcccctt agcttcttca
     2521 caattcagtt cccaacccta acaattcttg ttcatatcgc ttctctctac tttcaaggta
     2581 tgatccaatt tctctcttct ttctctgtaa tcctttcgtt gagttttgtt tccgatcaat
     2641 cataggtagt tttcttgttt cgaagcatga gatctaggaa ttttttgtga ttttccaaaa
     2701 ttgagatcgg tttgaaattg aattttacag cttgaatctc agatcttgtt ttatcaatgt
     2761 tttcgatggc ttgcgatgta gatctatgat aattgtggtt cagttttgtt aggaatcgat
     2821 ttcggtttag caattgcaga ttaattaggg tttccaattg aattcttcag atccgttatg
     2881 gaattatgtc aaataattta ttcaaattgg aaattattgt tagatccact cttaatctgt
     2941 ttgatccaag cttcaattag ggttttcact tgtttcaatt tcttgttatg gattctgatt
     3001 tatctgttga tgttagatcc actcttaatc tgtttgatcc aagcgttaat tagggttttc
     3061 acttgtttca atttcgtgtg ttggattctg atttatctgt tgttgatgtg attacagaat
     3121 ggcatttaaa gggtatatcc tatttctagt tatggtccta gtggccaata tgtgtactca
     3181 aaatgaagct caatcaggtt gtacatcaac gatcactagc ttgagccctt gtctaaacta
     3241 cataatgggg agttcttcaa atccatcatc ttcatgttgc tcacaacttt caagtgtagt
     3301 ccaatcttct cctcagtgtc tatgctccct ccttaatggt ggtggttcct cttttggaat
     3361 cactataaat caaacacttg ctctctctct cccttctgct tgtaaggttc aaactccacc
     3421 tgttagtcag tgcaaaggta tgcaagaata tgtttaatta taatagttgt tttagatcaa
     3481 tggtaaaatt gattgaatgt aaatacatgt atccacacta acatgtgtca gacagttgga
     3541 cacgtcttca acctcaagtg tcggtactaa tggaaaatta aactattaac ttattcccat
     3601 tgtataaatt caggtggcaa tggacagaca tctccaacaa gttcaacttc tccggcgggc
     3661 tctcctgttg attctccaac tgaatcaccc gaaggcgcaa taacaccttc tgctaactca
     3721 gattttccct caggtaaata tattattatg tttactaaag taattaatac atcaataatc
     3781 atactatagt atatagattt tattgattca ctcagttcac ctagtttttt cttaagcaat
     3841 ttagttgaag tagttttggt gaattgcagg tggagcagga tcaaaaagta taccatcaac
     3901 agatggtggc tcatccaatg gaagcactat tgaagtctcg ttcaacttgt ttctttcact
     3961 tcttgccatt gtgttttgtg tcatcaccaa gttctgagct tctctagcta gagtcgatcg
     4021 acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt
     4081 cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
     4141 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc
     4201 cagatcgcta agcttgaatt cttgcggaga gcggagaatt aagggagtca cgttatgacc
     4261 cccgccgatg acgcgggaca agccgtttta cgtttggaac tgacagaacc gcaacgttga
     4321 aggagccact cagccgcggg tttctggagt ttaatgagct aagcacatac gtcagaaacc
     4381 attattgcgc gttcaaaagt cgcctaaggt cactatcagc tagcaaatat ttcttgtcaa
     4441 aaatgctcca ctgacgttcc ataaattccc ctcggtatcc aattagagtc tcatattcac
     4501 tctcaaatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4561 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4621 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4681 gaactccaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4741 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4801 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4861 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4921 catcgcatcg agcgaggacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4981 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcggatg
     5041 cccgacggcg aggatctcgt cgtgacccag ggcgatgcct gcttgccgaa tatcatggtg
     5101 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5161 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5221 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5281 cttcttgacg agttcttctg agcttgtcaa gcagatcgtt caaacatttg gcaataaagt
     5341 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
     5401 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
     5461 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
     5521 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgacg ctggatccga
     5581 attcggcagg gcccactgca tccaccccag tacattaaaa acgtccgcaa tgtgttatta
     5641 agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc cagccaacag
     5701 ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc atcagccgtg
     5761 cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc
     5821 ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac
     5881 agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag
     5941 gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca
     6001 tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt
     6061 ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg
     6121 ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact
     6181 tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag
     6241 ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc
     6301 tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg
     6361 tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg
     6421 ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg
     6481 ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc
     6541 aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat
     6601 gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc
     6661 aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc
     6721 ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct
     6781 gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat
     6841 gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg
     6901 tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct
     6961 gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa
     7021 accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa
     7081 ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg
     7141 gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgtc
     7201 gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta
     7261 tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa
     7321 gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt
     7381 ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt
     7441 ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg
     7501 cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa
     7561 gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca
     7621 gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat
     7681 tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct
     7741 agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca
     7801 ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg
     7861 cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag
     7921 cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct
     7981 gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta
     8041 ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg
     8101 gcaaattgcc ctagcagggg aaaaaggtcg aaaaagcttc tttcctgtgg atagcacgta
     8161 cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc
     8221 gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt
     8281 ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata
     8341 actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg
     8401 ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc
     8461 tggcctacgg ccaggcaatc tac
//
