LOCUS       sequence-325320-        7641 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 7641)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7641)
  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            "unpublished"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:MtPSY1:T35s
FEATURES             Location/Qualifiers
     source          1..7641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..1207
                     /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      1273..1467
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1750..1854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1855..2649
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2820..3408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3309..3328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3531..3555
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        5074..5253
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5275..6069
                     /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(5329..5348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5939..5958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6082..6334
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6429..6453
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6717..7346
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt
       61 ggccaggcgt aagcggctgg gttgtctgcc ggccctgcaa tggcactgga acccccaagc
      121 ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct
      181 gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga
      241 ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc
      301 ccggcaaccg ccggcagccg gtgcgtcgtc gattaggaag ccgcccaagg gcgacgagca
      361 accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat
      421 ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta
      481 cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg
      541 ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg
      601 ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa
      661 gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg
      721 gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt
      781 gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg
      841 gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg
      901 caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg
      961 ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt
     1021 caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt
     1081 gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca
     1141 ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg
     1201 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
     1261 aagcttcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
     1321 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
     1381 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
     1441 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
     1501 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
     1561 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
     1621 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccg agctcccatc
     1681 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
     1741 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1801 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatggct
     1861 aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg cgcaaaagat
     1921 acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga aaacctatat
     1981 ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg ggaaaaggac
     2041 atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt tgaacggcat
     2101 gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc ggaagagtat
     2161 gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat caggctcttt
     2221 cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg cttagccgaa
     2281 ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg ggaagaggac
     2341 actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa gcccgaagag
     2401 gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa agatggcaaa
     2461 gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta tgacattgcc
     2521 ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga gctatttttt
     2581 gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt actggatgaa
     2641 ttgttttagc tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2701 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2761 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2821 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2881 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2941 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3001 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3061 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3121 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3181 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3241 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3301 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3361 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3421 gcggcctttt tacggttcct gtggttggca catacaaatg gacgaacgga taaacctttt
     3481 cacgcccttt taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc
     3541 caatatatcc tgtcaaacac tgatagttta aactccctcg gtcacatgtg catcctctag
     3601 tggagggaga gaggattttg aggaaataat taattgaatt acttgtatta ttgataaagt
     3661 aatttagata agttgttagt acaacttatt gcaactatgg atagacaaaa atcacttatt
     3721 ttaaggggag aagtatataa caacttatta taaaatttct gagcagccgg cctcctccaa
     3781 tcaatcatag agagtggagc ccattctgga aaaccaagga acccccaact tgcactggtg
     3841 cggtggccca attcaaaaaa aacggggccc agcaaagtaa cgccgttgta acgatttatc
     3901 aatccaaact caaaaggcgg cgagatgcgt ttaactccgt gaaattaaca aaccgccaac
     3961 aacttgcaat ttgcaactac cgtttccaag aagaactcaa ccacacaacg tatcctatcc
     4021 caaaccacac gcaactagtg acgcgtcata aggacacgtg tcacaatttg actggttaat
     4081 aatttcaccg cttttgctat aaattacctc caatcccctt agcttcttca caattcagtt
     4141 cccaacccta acaattcttg ttcatatcgc ttctctctac tttcaaggta tgatccaatt
     4201 tctctcttct ttctctgtaa tcctttcgtt gagttttgtt tccgatcaat cataggtagt
     4261 tttcttgttt cgaagcatga gatctaggaa ttttttgtga ttttccaaaa ttgagatcgg
     4321 tttgaaattg aattttacag cttgaatctc agatcttgtt ttatcaatgt tttcgatggc
     4381 ttgcgatgta gatctatgat aattgtggtt cagttttgtt aggaatcgat ttcggtttag
     4441 caattgcaga ttaattaggg tttccaattg aattcttcag atccgttatg gaattatgtc
     4501 aaataattta ttcaaattgg aaattattgt tagatccact cttaatctgt ttgatccaag
     4561 cttcaattag ggttttcact tgtttcaatt tcttgttatg gattctgatt tatctgttga
     4621 tgttagatcc actcttaatc tgtttgatcc aagcgttaat tagggttttc acttgtttca
     4681 atttcgtgtg ttggattctg atttatctgt tgttgatgtg attacagaat ggcattggct
     4741 tttgaagcac gaagtggctt gaccttctcc ttcttctttt tcttgctact atgctcagct
     4801 actgctcgga accctattat attttcaagt gatgaggctt taaacatggt tggaaggtct
     4861 ttgaaggtaa taaatcttga cgactatggt ggaccaactg ccaatcgtgg ccatgatcct
     4921 acattgaaca ggccggcaac cgggagaggt cgtcgtgtag ggagaaaagg atgagctaag
     4981 cttgaattct tgcggagagc ggagaattaa gggagtcacg ttatgacccc cgccgatgac
     5041 gcgggacaag ccgttttacg tttggaactg acagaaccgc aacgttgaag gagccactca
     5101 gccgcgggtt tctggagttt aatgagctaa gcacatacgt cagaaaccat tattgcgcgt
     5161 tcaaaagtcg cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa atgctccact
     5221 gacgttccat aaattcccct cggtatccaa ttagagtctc atattcactc tcaaatgatt
     5281 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5341 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5401 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actccaagac
     5461 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5521 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5581 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5641 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5701 cgaggacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5761 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcggatgcc cgacggcgag
     5821 gatctcgtcg tgacccaggg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5881 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5941 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6001 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6061 ttcttctgag cttgtcaagc agatcgttca aacatttggc aataaagttt cttaagattg
     6121 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat
     6181 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc
     6241 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
     6301 ttatcgcgcg cggtgtcatc tatgttacta gatcgacgct ggatccgaat tcggcagggc
     6361 ccactgcatc caccccagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc
     6421 gtcaatttgt ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg
     6481 cagctcggca caaaatcacc actcgataca ggcagcccat cagccgtgcg gctgcatgaa
     6541 atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa
     6601 gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat
     6661 gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga
     6721 aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc
     6781 tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg
     6841 gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg
     6901 accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg
     6961 acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc
     7021 tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg
     7081 ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg
     7141 cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc
     7201 ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca
     7261 caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg
     7321 ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac
     7381 aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag
     7441 cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac
     7501 caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata
     7561 catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg
     7621 ctaaaggagg cggcatggaa a
//
