LOCUS       sequence-325400-        8086 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 8086)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8086)
  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            "unpublished"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:MtMEG1:T35s
FEATURES             Location/Qualifiers
     source          1..8086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      73..267
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        550..654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             655..1449
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1620..2208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2109..2128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2331..2355
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    4045..4219
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        4319..4498
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4520..5314
                     /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(4574..4593)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5184..5203
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5327..5579
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5674..5698
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5962..6591
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(7020..8086,1..7)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
       61 aagcttcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
      121 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
      181 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
      241 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
      301 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
      361 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
      421 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccg agctcccatc
      481 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      541 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      601 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatggct
      661 aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg cgcaaaagat
      721 acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga aaacctatat
      781 ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg ggaaaaggac
      841 atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt tgaacggcat
      901 gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc ggaagagtat
      961 gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat caggctcttt
     1021 cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg cttagccgaa
     1081 ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg ggaagaggac
     1141 actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa gcccgaagag
     1201 gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa agatggcaaa
     1261 gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta tgacattgcc
     1321 ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga gctatttttt
     1381 gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt actggatgaa
     1441 ttgttttagc tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1981 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2221 gcggcctttt tacggttcct gtggttggca catacaaatg gacgaacgga taaacctttt
     2281 cacgcccttt taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc
     2341 caatatatcc tgtcaaacac tgatagttta aactccctcg gtcacatgtg catcctctag
     2401 tggagggaga gaggattttg aggaaataat taattgaatt acttgtatta ttgataaagt
     2461 aatttagata agttgttagt acaacttatt gcaactatgg atagacaaaa atcacttatt
     2521 ttaaggggag aagtatataa caacttatta taaaatttct gagcagccgg cctcctccaa
     2581 tcaatcatag agagtggagc ccattctgga aaaccaagga acccccaact tgcactggtg
     2641 cggtggccca attcaaaaaa aacggggccc agcaaagtaa cgccgttgta acgatttatc
     2701 aatccaaact caaaaggcgg cgagatgcgt ttaactccgt gaaattaaca aaccgccaac
     2761 aacttgcaat ttgcaactac cgtttccaag aagaactcaa ccacacaacg tatcctatcc
     2821 caaaccacac gcaactagtg acgcgtcata aggacacgtg tcacaatttg actggttaat
     2881 aatttcaccg cttttgctat aaattacctc caatcccctt agcttcttca caattcagtt
     2941 cccaacccta acaattcttg ttcatatcgc ttctctctac tttcaaggta tgatccaatt
     3001 tctctcttct ttctctgtaa tcctttcgtt gagttttgtt tccgatcaat cataggtagt
     3061 tttcttgttt cgaagcatga gatctaggaa ttttttgtga ttttccaaaa ttgagatcgg
     3121 tttgaaattg aattttacag cttgaatctc agatcttgtt ttatcaatgt tttcgatggc
     3181 ttgcgatgta gatctatgat aattgtggtt cagttttgtt aggaatcgat ttcggtttag
     3241 caattgcaga ttaattaggg tttccaattg aattcttcag atccgttatg gaattatgtc
     3301 aaataattta ttcaaattgg aaattattgt tagatccact cttaatctgt ttgatccaag
     3361 cttcaattag ggttttcact tgtttcaatt tcttgttatg gattctgatt tatctgttga
     3421 tgttagatcc actcttaatc tgtttgatcc aagcgttaat tagggttttc acttgtttca
     3481 atttcgtgtg ttggattctg atttatctgt tgttgatgtg attacagaat gaagctcttg
     3541 tctgttacaa aaccaaccat gatggttcct tctcattgtc tcatcttctc tctcattttt
     3601 gtttccattt tagccaatga agcaaccttg ctccatgtaa gtcctctctc tttctctgct
     3661 cataatatat gtataccctt ttgaatttaa ttcatggatc tagctgattt ttttttgtgg
     3721 gtagtgttaa aaattgaaaa ggattgtttt tttatgatta ggaagcaaat gggtcatttc
     3781 caatggtgcc tgtgatggaa gctgggaaga tgatgatgat gatgaatgag agcagaagaa
     3841 aacttggaag cttcaggata tgtgctatat gcacctgctg tggtggagca aaagggattt
     3901 gcatagcttc tccttgttgc tatgctatag attgcaacat tcccaataaa ccttttggtt
     3961 tatgttcatt cattcccaag gcttgtaatt gctttggatg ccatctttaa gcttctctag
     4021 ctagagtcga tcgacaagct cgagtttctc cataataatg tgtgagtagt tcccagataa
     4081 gggaattagg gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta
     4141 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     4201 cagtactaaa atccagatcg ctaagcttga attcttgcgg agagcggaga attaagggag
     4261 tcacgttatg acccccgccg atgacgcggg acaagccgtt ttacgtttgg aactgacaga
     4321 accgcaacgt tgaaggagcc actcagccgc gggtttctgg agtttaatga gctaagcaca
     4381 tacgtcagaa accattattg cgcgttcaaa agtcgcctaa ggtcactatc agctagcaaa
     4441 tatttcttgt caaaaatgct ccactgacgt tccataaatt cccctcggta tccaattaga
     4501 gtctcatatt cactctcaaa tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4561 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4621 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4681 cggtgccctg aatgaactcc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4741 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4801 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4861 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4921 ccaccaagcg aaacatcgca tcgagcgagg acgtactcgg atggaagccg gtcttgtcga
     4981 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5041 caaggcgcgg atgcccgacg gcgaggatct cgtcgtgacc cagggcgatg cctgcttgcc
     5101 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5161 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5221 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5281 cgccttctat cgccttcttg acgagttctt ctgagcttgt caagcagatc gttcaaacat
     5341 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
     5401 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat
     5461 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa
     5521 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg
     5581 acgctggatc cgaattcggc agggcccact gcatccaccc cagtacatta aaaacgtccg
     5641 caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc
     5701 agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag
     5761 cccatcagcc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca agctggcggc
     5821 ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt
     5881 atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa
     5941 acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca
     6001 ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt
     6061 ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat
     6121 caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc
     6181 ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc
     6241 gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacatatgg
     6301 gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga tggaaggcta
     6361 caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc
     6421 gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc
     6481 tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga gggcgacgct
     6541 gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg agttaatgag
     6601 gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg agcgcacgca
     6661 gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag cgggtcaact
     6721 ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc caaggcaaga
     6781 ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg agcaaatgaa
     6841 taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca agaacaacca
     6901 ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc
     6961 ggctgggttg tctgccggcc ctgcaatggc actggaaccc ccaagcccga ggaatcggcg
     7021 tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg
     7081 tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca gaagcacgcc
     7141 ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg caaccgccgg
     7201 cagccggtgc gtcgtcgatt aggaagccgc ccaagggcga cgagcaacca gattttttcg
     7261 ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac gtggccgttt
     7321 tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag cttccagacg
     7381 ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat tacgacctgg
     7441 tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa gggaagggag
     7501 acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc tgccggcgag
     7561 ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta aacaccacgc
     7621 acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg gtatccgagg
     7681 gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg ccggagtaca
     7741 tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag aacccggacg
     7801 tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt tttctctacc
     7861 gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag acgatctacg
     7921 aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc aagctgatcg
     7981 ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct ggcccgatcc
     8041 tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccgg
//
