LOCUS       sequence-324493-        7979 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 7979)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7979)
  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            "unpublished"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:MtCEP7:T35s
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      299..493
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        776..880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             881..1675
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1846..2434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2335..2354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2557..2581
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    4164..4338
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        4438..4617
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4639..5433
                     /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(4693..4712)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5303..5322
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5446..5698
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5793..5817
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6081..6710
                     /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(7139..7979,1..233)
                     /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 gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag acgatctacg
       61 aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc aagctgatcg
      121 ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct ggcccgatcc
      181 tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg
      241 agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaaagc ttctttcctg
      301 tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac ccgtacattg
      361 ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat ataaaagaga
      421 aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt aaaacccgcc
      481 tggcctgtgc ataactgtct ggccagcgca cagccgaaga gctgcaaaaa gcgcctaccc
      541 ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg gccgctggcc
      601 gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa gccgcgccgt
      661 cgccactcga ccgccggcgc ccacatcaag gcaccgagct cccatccccc caggggctgc
      721 gcccctcggc cgcgaacggc ctcaccccaa aaatggcagc gctggccaat tcgtgcgcgg
      781 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      841 accttgataa atgcttcaat aatattgaaa aaggaagagt atggctaaaa tgagaatatc
      901 accggaattg aaaaaactga tcgaaaaata ccgctgcgca aaagatacgg aaggaatgtc
      961 tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa aaatgacgga
     1021 cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga tgctatggct
     1081 ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg gctggagcaa
     1141 tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag atgaacaaag
     1201 ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact ccatcgacat
     1261 atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg attacttact
     1321 gaataacgat ctggccgatg tggattgcga aaactgggaa gaggacactc catttaaaga
     1381 tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc
     1441 ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat
     1501 tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct gcgtccggtc
     1561 gatcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact tactggggat
     1621 caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt tttagctgtc
     1681 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1741 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1801 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1861 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1921 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1981 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2041 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2101 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2161 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2221 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2281 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2341 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2401 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2461 gttcctgtgg ttggcacata caaatggacg aacggataaa ccttttcacg cccttttaaa
     2521 tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc
     2581 aaacactgat agtttaaact ccctcggtca catgtgcatc ctctagtgga gggagagagg
     2641 attttgagga aataattaat tgaattactt gtattattga taaagtaatt tagataagtt
     2701 gttagtacaa cttattgcaa ctatggatag acaaaaatca cttattttaa ggggagaagt
     2761 atataacaac ttattataaa atttctgagc agccggcctc ctccaatcaa tcatagagag
     2821 tggagcccat tctggaaaac caaggaaccc ccaacttgca ctggtgcggt ggcccaattc
     2881 aaaaaaaacg gggcccagca aagtaacgcc gttgtaacga tttatcaatc caaactcaaa
     2941 aggcggcgag atgcgtttaa ctccgtgaaa ttaacaaacc gccaacaact tgcaatttgc
     3001 aactaccgtt tccaagaaga actcaaccac acaacgtatc ctatcccaaa ccacacgcaa
     3061 ctagtgacgc gtcataagga cacgtgtcac aatttgactg gttaataatt tcaccgcttt
     3121 tgctataaat tacctccaat ccccttagct tcttcacaat tcagttccca accctaacaa
     3181 ttcttgttca tatcgcttct ctctactttc aaggtatgat ccaatttctc tcttctttct
     3241 ctgtaatcct ttcgttgagt tttgtttccg atcaatcata ggtagttttc ttgtttcgaa
     3301 gcatgagatc taggaatttt ttgtgatttt ccaaaattga gatcggtttg aaattgaatt
     3361 ttacagcttg aatctcagat cttgttttat caatgttttc gatggcttgc gatgtagatc
     3421 tatgataatt gtggttcagt tttgttagga atcgatttcg gtttagcaat tgcagattaa
     3481 ttagggtttc caattgaatt cttcagatcc gttatggaat tatgtcaaat aatttattca
     3541 aattggaaat tattgttaga tccactctta atctgtttga tccaagcttc aattagggtt
     3601 ttcacttgtt tcaatttctt gttatggatt ctgatttatc tgttgatgtt agatccactc
     3661 ttaatctgtt tgatccaagc gttaattagg gttttcactt gtttcaattt cgtgtgttgg
     3721 attctgattt atctgttgtt gatgtgatta cagaatgggt ttgtttcaag tcacaacaaa
     3781 atatttgatc gttattctag cactaagtat tgtatacaat tcctttcaaa taactcaagc
     3841 caggccaatt aaaccattga atcaacaatc ttcattaaac acacaagact cgggtgcaat
     3901 ccacactaac tcttttcggc cgacaacacc aggaagtagt cctggtgttg gccaccgaaa
     3961 ttttgttgta ggagataaga acacgagaac aatggtggtt gttcagagcc cggatgttga
     4021 ggtttttgtg acgaataaga gatccgatga tggtttcaaa cctacaaatc ctagtcatag
     4081 tcctggagtt ggccatggtt accataccaa aattagacat ttaaattagg cttctctagc
     4141 tagagtcgat cgacaagctc gagtttctcc ataataatgt gtgagtagtt cccagataag
     4201 ggaattaggg ttcctatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat
     4261 gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc
     4321 agtactaaaa tccagatcgc taagcttgaa ttcttgcgga gagcggagaa ttaagggagt
     4381 cacgttatga cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa
     4441 ccgcaacgtt gaaggagcca ctcagccgcg ggtttctgga gtttaatgag ctaagcacat
     4501 acgtcagaaa ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagcaaat
     4561 atttcttgtc aaaaatgctc cactgacgtt ccataaattc ccctcggtat ccaattagag
     4621 tctcatattc actctcaaat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4681 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4741 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4801 ggtgccctga atgaactcca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4861 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4921 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4981 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5041 caccaagcga aacatcgcat cgagcgagga cgtactcgga tggaagccgg tcttgtcgat
     5101 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5161 aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc agggcgatgc ctgcttgccg
     5221 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5281 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5341 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5401 gccttctatc gccttcttga cgagttcttc tgagcttgtc aagcagatcg ttcaaacatt
     5461 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     5521 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     5581 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     5641 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcga
     5701 cgctggatcc gaattcggca gggcccactg catccacccc agtacattaa aaacgtccgc
     5761 aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca
     5821 gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc
     5881 ccatcagccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc
     5941 tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta
     6001 tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa
     6061 caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag
     6121 gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc
     6181 tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc
     6241 aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg
     6301 gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg
     6361 cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg
     6421 ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac
     6481 aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg
     6541 aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct
     6601 acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg
     6661 cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg
     6721 taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag
     6781 cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt
     6841 tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac
     6901 cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat
     6961 aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag
     7021 gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg
     7081 gctgggttgt ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt
     7141 gagcggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt
     7201 ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc
     7261 cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc
     7321 agccggtgcg tcgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt
     7381 tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt
     7441 ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg
     7501 gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt
     7561 actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga
     7621 caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc
     7681 cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca
     7741 cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg
     7801 tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat
     7861 cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt
     7921 gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt ttctctacc
//
