LOCUS       sequence-325438-        7988 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 7988)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7988)
  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            "unpublished"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:MtFam208_1:T35s
FEATURES             Location/Qualifiers
     source          1..7988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             190..1263
                     /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      1329..1523
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1806..1910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1911..2705
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2876..3464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3365..3384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3587..3611
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    5203..5377
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5477..5656
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5678..6472
                     /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(5732..5751)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6342..6361
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6485..6737
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6832..6856
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7120..7749
                     /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 agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca
       61 agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc
      121 aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc actggaaccc ccaagcccga
      181 ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt
      241 gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca
      301 gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg
      361 caaccgccgg cagccggtgc gtcgtcgatt aggaagccgc ccaagggcga cgagcaacca
      421 gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac
      481 gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag
      541 cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat
      601 tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa
      661 gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc
      721 tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta
      781 aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg
      841 gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg
      901 ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag
      961 aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt
     1021 tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag
     1081 acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc
     1141 aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct
     1201 ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc
     1261 taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaaagc
     1321 ttctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac
     1381 ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat
     1441 ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt
     1501 aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga gctgcaaaaa
     1561 gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg
     1621 gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa
     1681 gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccgagct cccatccccc
     1741 caggggctgc gcccctcggc cgcgaacggc ctcaccccaa aaatggcagc gctggccaat
     1801 tcgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1861 tgagacaata accttgataa atgcttcaat aatattgaaa aaggaagagt atggctaaaa
     1921 tgagaatatc accggaattg aaaaaactga tcgaaaaata ccgctgcgca aaagatacgg
     1981 aaggaatgtc tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa
     2041 aaatgacgga cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga
     2101 tgctatggct ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg
     2161 gctggagcaa tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag
     2221 atgaacaaag ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact
     2281 ccatcgacat atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg
     2341 attacttact gaataacgat ctggccgatg tggattgcga aaactgggaa gaggacactc
     2401 catttaaaga tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac
     2461 ttgtcttttc ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa
     2521 gtggctttat tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct
     2581 gcgtccggtc gatcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact
     2641 tactggggat caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt
     2701 tttagctgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2761 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2821 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2881 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2941 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3001 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3061 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3121 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3181 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3241 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3301 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3361 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3421 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3481 cctttttacg gttcctgtgg ttggcacata caaatggacg aacggataaa ccttttcacg
     3541 cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat
     3601 atatcctgtc aaacactgat agtttaaact ccctcggtca catgtgcatc ctctagtgga
     3661 gggagagagg attttgagga aataattaat tgaattactt gtattattga taaagtaatt
     3721 tagataagtt gttagtacaa cttattgcaa ctatggatag acaaaaatca cttattttaa
     3781 ggggagaagt atataacaac ttattataaa atttctgagc agccggcctc ctccaatcaa
     3841 tcatagagag tggagcccat tctggaaaac caaggaaccc ccaacttgca ctggtgcggt
     3901 ggcccaattc aaaaaaaacg gggcccagca aagtaacgcc gttgtaacga tttatcaatc
     3961 caaactcaaa aggcggcgag atgcgtttaa ctccgtgaaa ttaacaaacc gccaacaact
     4021 tgcaatttgc aactaccgtt tccaagaaga actcaaccac acaacgtatc ctatcccaaa
     4081 ccacacgcaa ctagtgacgc gtcataagga cacgtgtcac aatttgactg gttaataatt
     4141 tcaccgcttt tgctataaat tacctccaat ccccttagct tcttcacaat tcagttccca
     4201 accctaacaa ttcttgttca tatcgcttct ctctactttc aaggtatgat ccaatttctc
     4261 tcttctttct ctgtaatcct ttcgttgagt tttgtttccg atcaatcata ggtagttttc
     4321 ttgtttcgaa gcatgagatc taggaatttt ttgtgatttt ccaaaattga gatcggtttg
     4381 aaattgaatt ttacagcttg aatctcagat cttgttttat caatgttttc gatggcttgc
     4441 gatgtagatc tatgataatt gtggttcagt tttgttagga atcgatttcg gtttagcaat
     4501 tgcagattaa ttagggtttc caattgaatt cttcagatcc gttatggaat tatgtcaaat
     4561 aatttattca aattggaaat tattgttaga tccactctta atctgtttga tccaagcttc
     4621 aattagggtt ttcacttgtt tcaatttctt gttatggatt ctgatttatc tgttgatgtt
     4681 agatccactc ttaatctgtt tgatccaagc gttaattagg gttttcactt gtttcaattt
     4741 cgtgtgttgg attctgattt atctgttgtt gatgtgatta cagaatgaaa atatccataa
     4801 tttcttctgt ggtgttaaca acactcctag tagcttcact ctttcaacca catcaacata
     4861 tctcagcagc accacagaac attactgatc aaggagctaa tggatgcatt gggcttcact
     4921 gcctaattgc cgtggacgat gaagccgatt tcttaatgga tcagcgtgct acaagaccga
     4981 gtagaatgct tgcaaatgct tttaactaca acatagcaaa aacaaatgat gctaataaac
     5041 aatcgcctga ttgtggagta atgggattga atggtcaaac tacatgtctt ggctccaaac
     5101 agctcgtaaa taaacaagga aggccttgcg accctacaaa cagagtgtat ccatactgca
     5161 cagcataggc ttctctagct agagtcgatc gacaagctcg agtttctcca taataatgtg
     5221 tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca
     5281 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta
     5341 attcctaaaa ccaaaatcca gtactaaaat ccagatcgct aagcttgaat tcttgcggag
     5401 agcggagaat taagggagtc acgttatgac ccccgccgat gacgcgggac aagccgtttt
     5461 acgtttggaa ctgacagaac cgcaacgttg aaggagccac tcagccgcgg gtttctggag
     5521 tttaatgagc taagcacata cgtcagaaac cattattgcg cgttcaaaag tcgcctaagg
     5581 tcactatcag ctagcaaata tttcttgtca aaaatgctcc actgacgttc cataaattcc
     5641 cctcggtatc caattagagt ctcatattca ctctcaaatg attgaacaag atggattgca
     5701 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5761 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5821 tgtcaagacc gacctgtccg gtgccctgaa tgaactccaa gacgaggcag cgcggctatc
     5881 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5941 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6001 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6061 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgaggac gtactcggat
     6121 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6181 cgaactgttc gccaggctca aggcgcggat gcccgacggc gaggatctcg tcgtgaccca
     6241 gggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6301 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6361 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6421 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcttgtca
     6481 agcagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     6541 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     6601 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     6661 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     6721 atctatgtta ctagatcgac gctggatccg aattcggcag ggcccactgc atccacccca
     6781 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
     6841 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
     6901 accactcgat acaggcagcc catcagccgt gcggctgcat gaaatcctgg ccggtttgtc
     6961 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     7021 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     7081 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     7141 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
     7201 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     7261 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     7321 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     7381 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     7441 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     7501 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     7561 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     7621 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     7681 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     7741 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     7801 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     7861 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     7921 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     7981 agtaaatg
//
