LOCUS       sequence-325454-        7835 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 7835)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7835)
  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            "unpublished"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:MtIDA6:T35s
FEATURES             Location/Qualifiers
     source          1..7835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..1244
                     /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      1310..1504
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1787..1891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1892..2686
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2857..3445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3346..3365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3568..3592
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    5031..5205
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5305..5484
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5506..6300
                     /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(5560..5579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6170..6189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6313..6565
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6660..6684
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6948..7577
                     /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 agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac
       61 gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag cggctgggtt
      121 gtctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgagcggtc
      181 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
      241 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
      301 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
      361 cgtcgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
      421 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
      481 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
      541 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
      601 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
      661 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
      721 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
      781 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
      841 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
      901 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
      961 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
     1021 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
     1081 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
     1141 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
     1201 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
     1261 tagggcaaat tgccctagca ggggaaaaag gtcgaaaaag cttctttcct gtggatagca
     1321 cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa
     1381 agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg
     1441 atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg
     1501 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
     1561 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
     1621 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
     1681 accgccggcg cccacatcaa ggcaccgagc tcccatcccc ccaggggctg cgcccctcgg
     1741 ccgcgaacgg cctcacccca aaaatggcag cgctggccaa ttcgtgcgcg gaacccctat
     1801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccttgata
     1861 aatgcttcaa taatattgaa aaaggaagag tatggctaaa atgagaatat caccggaatt
     1921 gaaaaaactg atcgaaaaat accgctgcgc aaaagatacg gaaggaatgt ctcctgctaa
     1981 ggtatataag ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta
     2041 taaagggacc acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa
     2101 gctgcctgtt ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat
     2161 gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa
     2221 gattatcgag ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg
     2281 tccctatacg aatagcttag acagccgctt agccgaattg gattacttac tgaataacga
     2341 tctggccgat gtggattgcg aaaactggga agaggacact ccatttaaag atccgcgcga
     2401 gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga
     2461 cctgggagac agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg
     2521 gagaagcggc agggcggaca agtggtatga cattgccttc tgcgtccggt cgatcaggga
     2581 ggatatcggg gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga
     2641 ttgggagaaa ataaaatatt atattttact ggatgaattg ttttagctgt cagaccaagt
     2701 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2761 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2821 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2881 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2941 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3001 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3061 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3121 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3181 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3241 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3301 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3361 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3421 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgtg
     3481 gttggcacat acaaatggac gaacggataa accttttcac gcccttttaa atatccgatt
     3541 attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga
     3601 tagtttaaac tccctcggtc acatgtgcat cctctagtgg agggagagag gattttgagg
     3661 aaataattaa ttgaattact tgtattattg ataaagtaat ttagataagt tgttagtaca
     3721 acttattgca actatggata gacaaaaatc acttatttta aggggagaag tatataacaa
     3781 cttattataa aatttctgag cagccggcct cctccaatca atcatagaga gtggagccca
     3841 ttctggaaaa ccaaggaacc cccaacttgc actggtgcgg tggcccaatt caaaaaaaac
     3901 ggggcccagc aaagtaacgc cgttgtaacg atttatcaat ccaaactcaa aaggcggcga
     3961 gatgcgttta actccgtgaa attaacaaac cgccaacaac ttgcaatttg caactaccgt
     4021 ttccaagaag aactcaacca cacaacgtat cctatcccaa accacacgca actagtgacg
     4081 cgtcataagg acacgtgtca caatttgact ggttaataat ttcaccgctt ttgctataaa
     4141 ttacctccaa tccccttagc ttcttcacaa ttcagttccc aaccctaaca attcttgttc
     4201 atatcgcttc tctctacttt caaggtatga tccaatttct ctcttctttc tctgtaatcc
     4261 tttcgttgag ttttgtttcc gatcaatcat aggtagtttt cttgtttcga agcatgagat
     4321 ctaggaattt tttgtgattt tccaaaattg agatcggttt gaaattgaat tttacagctt
     4381 gaatctcaga tcttgtttta tcaatgtttt cgatggcttg cgatgtagat ctatgataat
     4441 tgtggttcag ttttgttagg aatcgatttc ggtttagcaa ttgcagatta attagggttt
     4501 ccaattgaat tcttcagatc cgttatggaa ttatgtcaaa taatttattc aaattggaaa
     4561 ttattgttag atccactctt aatctgtttg atccaagctt caattagggt tttcacttgt
     4621 ttcaatttct tgttatggat tctgatttat ctgttgatgt tagatccact cttaatctgt
     4681 ttgatccaag cgttaattag ggttttcact tgtttcaatt tcgtgtgttg gattctgatt
     4741 tatctgttgt tgatgtgatt acagaatgaa taggatgcat ttcaatgtgg tactagtgct
     4801 tctacttttg gtgatatatt ttgttggtca tggttatggc tctagacaca atcaagtttt
     4861 caaagtttat cccaaaacta aggattttcc ttcaagtgtc cttggtttgt tgccaaaaac
     4921 aatgccaatt ccaccttccg ggccttctaa gaagcacaat gacattggaa tacaaagctc
     4981 aatgggaaaa ccataggctt ctctagctag agtcgatcga caagctcgag tttctccata
     5041 ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt
     5101 gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa
     5161 aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcgctaa gcttgaattc
     5221 ttgcggagag cggagaatta agggagtcac gttatgaccc ccgccgatga cgcgggacaa
     5281 gccgttttac gtttggaact gacagaaccg caacgttgaa ggagccactc agccgcgggt
     5341 ttctggagtt taatgagcta agcacatacg tcagaaacca ttattgcgcg ttcaaaagtc
     5401 gcctaaggtc actatcagct agcaaatatt tcttgtcaaa aatgctccac tgacgttcca
     5461 taaattcccc tcggtatcca attagagtct catattcact ctcaaatgat tgaacaagat
     5521 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5581 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5641 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactccaaga cgaggcagcg
     5701 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5761 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5821 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5881 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgaggacgt
     5941 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6001 gcgccagccg aactgttcgc caggctcaag gcgcggatgc ccgacggcga ggatctcgtc
     6061 gtgacccagg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6121 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6181 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6241 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6301 gcttgtcaag cagatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
     6361 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     6421 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     6481 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     6541 gcggtgtcat ctatgttact agatcgacgc tggatccgaa ttcggcaggg cccactgcat
     6601 ccaccccagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg
     6661 tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg gcagctcggc
     6721 acaaaatcac cactcgatac aggcagccca tcagccgtgc ggctgcatga aatcctggcc
     6781 ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag
     6841 cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct
     6901 gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg
     6961 ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg
     7021 ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc
     7081 gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga
     7141 cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc
     7201 cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg
     7261 tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc
     7321 gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag
     7381 gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg
     7441 agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc
     7501 ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta
     7561 aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct
     7621 aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga
     7681 cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa
     7741 gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca
     7801 gctaccagag taaatgagca aatgaataaa tgagt
//
