LOCUS       sequence-325396-        8012 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 8012)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8012)
  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            "unpublished"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:MtSTIG15:T35s
FEATURES             Location/Qualifiers
     source          1..8012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             157..1230
                     /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      1296..1490
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1773..1877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1878..2672
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2843..3431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3332..3351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3554..3578
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    5194..5368
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5468..5647
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5669..6463
                     /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(5723..5742)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6333..6352
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6476..6728
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6823..6847
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7111..7740
                     /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 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
       61 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct gccggccctg
      121 caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc
      181 ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag
      241 gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc
      301 gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgtc gtcgattagg
      361 aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc
      421 acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga
      481 cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg
      541 ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta
      601 accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt
      661 ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac
      721 gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag
      781 aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac
      841 aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct agctgattgg
      901 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac
      961 tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc
     1021 aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag
     1081 ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac
     1141 gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg
     1201 atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc
     1261 ctagcagggg aaaaaggtcg aaaaagcttc tttcctgtgg atagcacgta cattgggaac
     1321 ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc gtacattggg
     1381 aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt ttccgcctaa
     1441 aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata actgtctggc
     1501 cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc
     1561 cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc tggcctacgg
     1621 ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca
     1681 catcaaggca ccgagctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc
     1741 accccaaaaa tggcagcgct ggccaattcg tgcgcggaac ccctatttgt ttatttttct
     1801 aaatacattc aaatatgtat ccgctcatga gacaataacc ttgataaatg cttcaataat
     1861 attgaaaaag gaagagtatg gctaaaatga gaatatcacc ggaattgaaa aaactgatcg
     1921 aaaaataccg ctgcgcaaaa gatacggaag gaatgtctcc tgctaaggta tataagctgg
     1981 tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa gggaccacct
     2041 atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg cctgttccaa
     2101 aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt gaggccgatg
     2161 gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt atcgagctgt
     2221 atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc tatacgaata
     2281 gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg gccgatgtgg
     2341 attgcgaaaa ctgggaagag gacactccat ttaaagatcc gcgcgagctg tatgattttt
     2401 taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg ggagacagca
     2461 acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga agcggcaggg
     2521 cggacaagtg gtatgacatt gccttctgcg tccggtcgat cagggaggat atcggggaag
     2581 aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg gagaaaataa
     2641 aatattatat tttactggat gaattgtttt agctgtcaga ccaagtttac tcatatatac
     2701 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2761 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctgtggttg gcacatacaa
     3481 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
     3541 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaactccc
     3601 tcggtcacat gtgcatcctc tagtggaggg agagaggatt ttgaggaaat aattaattga
     3661 attacttgta ttattgataa agtaatttag ataagttgtt agtacaactt attgcaacta
     3721 tggatagaca aaaatcactt attttaaggg gagaagtata taacaactta ttataaaatt
     3781 tctgagcagc cggcctcctc caatcaatca tagagagtgg agcccattct ggaaaaccaa
     3841 ggaaccccca acttgcactg gtgcggtggc ccaattcaaa aaaaacgggg cccagcaaag
     3901 taacgccgtt gtaacgattt atcaatccaa actcaaaagg cggcgagatg cgtttaactc
     3961 cgtgaaatta acaaaccgcc aacaacttgc aatttgcaac taccgtttcc aagaagaact
     4021 caaccacaca acgtatccta tcccaaacca cacgcaacta gtgacgcgtc ataaggacac
     4081 gtgtcacaat ttgactggtt aataatttca ccgcttttgc tataaattac ctccaatccc
     4141 cttagcttct tcacaattca gttcccaacc ctaacaattc ttgttcatat cgcttctctc
     4201 tactttcaag gtatgatcca atttctctct tctttctctg taatcctttc gttgagtttt
     4261 gtttccgatc aatcataggt agttttcttg tttcgaagca tgagatctag gaattttttg
     4321 tgattttcca aaattgagat cggtttgaaa ttgaatttta cagcttgaat ctcagatctt
     4381 gttttatcaa tgttttcgat ggcttgcgat gtagatctat gataattgtg gttcagtttt
     4441 gttaggaatc gatttcggtt tagcaattgc agattaatta gggtttccaa ttgaattctt
     4501 cagatccgtt atggaattat gtcaaataat ttattcaaat tggaaattat tgttagatcc
     4561 actcttaatc tgtttgatcc aagcttcaat tagggttttc acttgtttca atttcttgtt
     4621 atggattctg atttatctgt tgatgttaga tccactctta atctgtttga tccaagcgtt
     4681 aattagggtt ttcacttgtt tcaatttcgt gtgttggatt ctgatttatc tgttgttgat
     4741 gtgattacag aatgaagtcc atgaagactc ttttccttgt gaccttgtta attatggctt
     4801 tgaccgttac accatcatca gcatcatctt ctgaaaacga agaaccaaac tcttcacttc
     4861 aaggaacaag ccacttcctc aaccgcaaac aatatagaat ctctctcaca tgtgacaaat
     4921 atcccaagat ttgccatacc aaaggtagcg caggtcctga ttgttgcaac aacaaatgtg
     4981 tcaattttac catagacatg tttaactgtg ggaggtgtgg aaagaaatgc agtttcccaa
     5041 agatatgctg tgaaggtaaa tgtgtgaacc ctaggagtaa caagaaacat tgtgggaaat
     5101 gtggtaacaa gtgtgaaagt agaggttctt gtgtttatgg aatgtgcagc tatgcatagg
     5161 cttctctagc tagagtcgat cgacaagctc gagtttctcc ataataatgt gtgagtagtt
     5221 cccagataag ggaattaggg ttcctatagg gtttcgctca tgtgttgagc atataagaaa
     5281 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     5341 accaaaatcc agtactaaaa tccagatcgc taagcttgaa ttcttgcgga gagcggagaa
     5401 ttaagggagt cacgttatga cccccgccga tgacgcggga caagccgttt tacgtttgga
     5461 actgacagaa ccgcaacgtt gaaggagcca ctcagccgcg ggtttctgga gtttaatgag
     5521 ctaagcacat acgtcagaaa ccattattgc gcgttcaaaa gtcgcctaag gtcactatca
     5581 gctagcaaat atttcttgtc aaaaatgctc cactgacgtt ccataaattc ccctcggtat
     5641 ccaattagag tctcatattc actctcaaat gattgaacaa gatggattgc acgcaggttc
     5701 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5761 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5821 cgacctgtcc ggtgccctga atgaactcca agacgaggca gcgcggctat cgtggctggc
     5881 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5941 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6001 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6061 cccattcgac caccaagcga aacatcgcat cgagcgagga cgtactcgga tggaagccgg
     6121 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6181 cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc agggcgatgc
     6241 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6301 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6361 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6421 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcttgtc aagcagatcg
     6481 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     6541 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     6601 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     6661 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     6721 actagatcga cgctggatcc gaattcggca gggcccactg catccacccc agtacattaa
     6781 aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat
     6841 cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga
     6901 tacaggcagc ccatcagccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa
     6961 gctggcggcc tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag
     7021 gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg
     7081 agtaaataaa caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa
     7141 ggcgggtcag gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg
     7201 gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg
     7261 cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg
     7321 aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg
     7381 gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac
     7441 gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat
     7501 ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt
     7561 gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag
     7621 cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag
     7681 ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga
     7741 gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga
     7801 gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc
     7861 gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc
     7921 aaggcaagac cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga
     7981 gcaaatgaat aaatgagtag atgaatttta gc
//
