LOCUS       sequence-324495-        8060 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 8060)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8060)
  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            "unpublished"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:MtCEP9:T35s
FEATURES             Location/Qualifiers
     source          1..8060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             173..1246
                     /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      1312..1506
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1789..1893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1894..2688
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2859..3447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3348..3367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3570..3594
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    5258..5432
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5532..5711
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5733..6527
                     /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(5787..5806)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6397..6416
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6540..6792
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6887..6911
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7175..7804
                     /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 gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg
       61 acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg
      121 ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg
      181 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
      241 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
      301 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
      361 tgcgtcgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
      421 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
      481 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
      541 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
      601 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
      661 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
      721 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
      781 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
      841 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
      901 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
      961 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
     1021 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
     1081 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
     1141 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat
     1201 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat
     1261 gctagggcaa attgccctag caggggaaaa aggtcgaaaa agcttctttc ctgtggatag
     1321 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc
     1381 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg
     1441 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg
     1501 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc
     1561 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa
     1621 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact
     1681 cgaccgccgg cgcccacatc aaggcaccga gctcccatcc ccccaggggc tgcgcccctc
     1741 ggccgcgaac ggcctcaccc caaaaatggc agcgctggcc aattcgtgcg cggaacccct
     1801 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccttga
     1861 taaatgcttc aataatattg aaaaaggaag agtatggcta aaatgagaat atcaccggaa
     1921 ttgaaaaaac tgatcgaaaa ataccgctgc gcaaaagata cggaaggaat gtctcctgct
     1981 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
     2041 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
     2101 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
     2161 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa
     2221 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
     2281 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
     2341 gatctggccg atgtggattg cgaaaactgg gaagaggaca ctccatttaa agatccgcgc
     2401 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
     2461 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
     2521 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
     2581 gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
     2641 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagct gtcagaccaa
     2701 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2761 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2821 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2881 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2941 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3001 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3061 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3121 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3181 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3241 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3301 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3361 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3421 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3481 tggttggcac atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga
     3541 ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcaaacact
     3601 gatagtttaa actccctcgg tcacatgtgc atcctctagt ggagggagag aggattttga
     3661 ggaaataatt aattgaatta cttgtattat tgataaagta atttagataa gttgttagta
     3721 caacttattg caactatgga tagacaaaaa tcacttattt taaggggaga agtatataac
     3781 aacttattat aaaatttctg agcagccggc ctcctccaat caatcataga gagtggagcc
     3841 cattctggaa aaccaaggaa cccccaactt gcactggtgc ggtggcccaa ttcaaaaaaa
     3901 acggggccca gcaaagtaac gccgttgtaa cgatttatca atccaaactc aaaaggcggc
     3961 gagatgcgtt taactccgtg aaattaacaa accgccaaca acttgcaatt tgcaactacc
     4021 gtttccaaga agaactcaac cacacaacgt atcctatccc aaaccacacg caactagtga
     4081 cgcgtcataa ggacacgtgt cacaatttga ctggttaata atttcaccgc ttttgctata
     4141 aattacctcc aatcccctta gcttcttcac aattcagttc ccaaccctaa caattcttgt
     4201 tcatatcgct tctctctact ttcaaggtat gatccaattt ctctcttctt tctctgtaat
     4261 cctttcgttg agttttgttt ccgatcaatc ataggtagtt ttcttgtttc gaagcatgag
     4321 atctaggaat tttttgtgat tttccaaaat tgagatcggt ttgaaattga attttacagc
     4381 ttgaatctca gatcttgttt tatcaatgtt ttcgatggct tgcgatgtag atctatgata
     4441 attgtggttc agttttgtta ggaatcgatt tcggtttagc aattgcagat taattagggt
     4501 ttccaattga attcttcaga tccgttatgg aattatgtca aataatttat tcaaattgga
     4561 aattattgtt agatccactc ttaatctgtt tgatccaagc ttcaattagg gttttcactt
     4621 gtttcaattt cttgttatgg attctgattt atctgttgat gttagatcca ctcttaatct
     4681 gtttgatcca agcgttaatt agggttttca cttgtttcaa tttcgtgtgt tggattctga
     4741 tttatctgtt gttgatgtga ttacagaatg ggtgaatttc aggccatgca aaaatatttt
     4801 gccatttttc ttgtattagt tgcctaccat attttccttc caactcaagc taggaagata
     4861 aaaccattga ttgaagataa tcccaaacct accttcacat cccttaaaac tgctgtaaat
     4921 attccttctc caacatttga gaagaaagtt aaccttccca tgatgccaaa tcatggtgtc
     4981 gcaagtatag gagattcaag cggagataca aatgctttcc gacccacaac accaggaagc
     5041 agtcctggtg ttggtcatcg gaagtttgta ggagaggtta aagatagtac agttgttcgg
     5101 agtccgaatg ttaaagtttt tgtgacttct gagagatcaa aagatgcttt taaacctact
     5161 tacccaaatc atagcccagg tgttggacat gttaaccaaa gcacaaaagg acaactaaat
     5221 taggcttctc tagctagagt cgatcgacaa gctcgagttt ctccataata atgtgtgagt
     5281 agttcccaga taagggaatt agggttccta tagggtttcg ctcatgtgtt gagcatataa
     5341 gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc
     5401 taaaaccaaa atccagtact aaaatccaga tcgctaagct tgaattcttg cggagagcgg
     5461 agaattaagg gagtcacgtt atgacccccg ccgatgacgc gggacaagcc gttttacgtt
     5521 tggaactgac agaaccgcaa cgttgaagga gccactcagc cgcgggtttc tggagtttaa
     5581 tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact
     5641 atcagctagc aaatatttct tgtcaaaaat gctccactga cgttccataa attcccctcg
     5701 gtatccaatt agagtctcat attcactctc aaatgattga acaagatgga ttgcacgcag
     5761 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5821 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5881 agaccgacct gtccggtgcc ctgaatgaac tccaagacga ggcagcgcgg ctatcgtggc
     5941 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6001 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6061 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6121 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg aggacgtact cggatggaag
     6181 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6241 tgttcgccag gctcaaggcg cggatgcccg acggcgagga tctcgtcgtg acccagggcg
     6301 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6361 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6421 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6481 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagct tgtcaagcag
     6541 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga
     6601 tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca
     6661 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg
     6721 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta
     6781 tgttactaga tcgacgctgg atccgaattc ggcagggccc actgcatcca ccccagtaca
     6841 ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat
     6901 atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac
     6961 tcgatacagg cagcccatca gccgtgcggc tgcatgaaat cctggccggt ttgtctgatg
     7021 ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa
     7081 aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc
     7141 gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca
     7201 gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc
     7261 cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc
     7321 cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga
     7381 cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga
     7441 cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc
     7501 ttacgacata tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac
     7561 ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg
     7621 cggtgaggtt gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac
     7681 gcagcgcgtg agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc
     7741 cgagggcgac gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat
     7801 ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt
     7861 ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg
     7921 aagcgggtca actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta
     7981 cgccaaggca agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa
     8041 atgagcaaat gaataaatga
//
