LOCUS       sequence-324761-        8009 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 8009)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8009)
  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            "unpublished"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:MtSTIG8:T35s
FEATURES             Location/Qualifiers
     source          1..8009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..206
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             228..1022
                     /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(282..301)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     892..911
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1035..1287
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1382..1406
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             1670..2299
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2728..3801
                     /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      3867..4061
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4344..4448
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4449..5243
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      5414..6002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5903..5922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6125..6149
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    7762..7936
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 aagccgtttt acgtttggaa ctgacagaac cgcaacgttg aaggagccac tcagccgcgg
       61 gtttctggag tttaatgagc taagcacata cgtcagaaac cattattgcg cgttcaaaag
      121 tcgcctaagg tcactatcag ctagcaaata tttcttgtca aaaatgctcc actgacgttc
      181 cataaattcc cctcggtatc caattagagt ctcatattca ctctcaaatg attgaacaag
      241 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
      301 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
      361 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactccaa gacgaggcag
      421 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
      481 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
      541 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
      601 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgaggac
      661 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
      721 tcgcgccagc cgaactgttc gccaggctca aggcgcggat gcccgacggc gaggatctcg
      781 tcgtgaccca gggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
      841 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
      901 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
      961 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     1021 gagcttgtca agcagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg
     1081 ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa
     1141 ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat
     1201 tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc
     1261 gcgcggtgtc atctatgtta ctagatcgac gctggatccg aattcggcag ggcccactgc
     1321 atccacccca gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt
     1381 tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg
     1441 gcacaaaatc accactcgat acaggcagcc catcagccgt gcggctgcat gaaatcctgg
     1501 ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg
     1561 agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg
     1621 ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat
     1681 cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg
     1741 cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc
     1801 ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc
     1861 gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc
     1921 gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc
     1981 ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca
     2041 gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa
     2101 aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct
     2161 tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt
     2221 tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat
     2281 taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg
     2341 ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca
     2401 gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg
     2461 aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg
     2521 cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg
     2581 aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg
     2641 aggaacgggc ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac
     2701 tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa
     2761 atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag
     2821 cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga
     2881 atccgcaaag aatcccggca accgccggca gccggtgcgt cgtcgattag gaagccgccc
     2941 aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat
     3001 agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc
     3061 gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc
     3121 atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc
     3181 atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt
     3241 gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta
     3301 gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag
     3361 aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta
     3421 aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc
     3481 gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc
     3541 gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa
     3601 gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag
     3661 ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag
     3721 gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc
     3781 gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg
     3841 gaaaaaggtc gaaaaagctt ctttcctgtg gatagcacgt acattgggaa cccaaagccg
     3901 tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca
     3961 cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta
     4021 aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca
     4081 gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct
     4141 tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat
     4201 ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc
     4261 accgagctcc catcccccca ggggctgcgc ccctcggccg cgaacggcct caccccaaaa
     4321 atggcagcgc tggccaattc gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4381 caaatatgta tccgctcatg agacaataac cttgataaat gcttcaataa tattgaaaaa
     4441 ggaagagtat ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc
     4501 gctgcgcaaa agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa
     4561 atgaaaacct atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg
     4621 aacgggaaaa ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc
     4681 actttgaacg gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt
     4741 gctcggaaga gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt
     4801 gcatcaggct ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca
     4861 gccgcttagc cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa
     4921 actgggaaga ggacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg
     4981 aaaagcccga agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg
     5041 tgaaagatgg caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt
     5101 ggtatgacat tgccttctgc gtccggtcga tcagggagga tatcggggaa gaacagtatg
     5161 tcgagctatt ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata
     5221 ttttactgga tgaattgttt tagctgtcag accaagttta ctcatatata ctttagattg
     5281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5521 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6001 aaaacgccag caacgcggcc tttttacggt tcctgtggtt ggcacataca aatggacgaa
     6061 cggataaacc ttttcacgcc cttttaaata tccgattatt ctaataaacg ctcttttctc
     6121 ttaggtttac ccgccaatat atcctgtcaa acactgatag tttaaactcc ctcggtcaca
     6181 tgtgcatcct ctagtggagg gagagaggat tttgaggaaa taattaattg aattacttgt
     6241 attattgata aagtaattta gataagttgt tagtacaact tattgcaact atggatagac
     6301 aaaaatcact tattttaagg ggagaagtat ataacaactt attataaaat ttctgagcag
     6361 ccggcctcct ccaatcaatc atagagagtg gagcccattc tggaaaacca aggaaccccc
     6421 aacttgcact ggtgcggtgg cccaattcaa aaaaaacggg gcccagcaaa gtaacgccgt
     6481 tgtaacgatt tatcaatcca aactcaaaag gcggcgagat gcgtttaact ccgtgaaatt
     6541 aacaaaccgc caacaacttg caatttgcaa ctaccgtttc caagaagaac tcaaccacac
     6601 aacgtatcct atcccaaacc acacgcaact agtgacgcgt cataaggaca cgtgtcacaa
     6661 tttgactggt taataatttc accgcttttg ctataaatta cctccaatcc ccttagcttc
     6721 ttcacaattc agttcccaac cctaacaatt cttgttcata tcgcttctct ctactttcaa
     6781 ggtatgatcc aatttctctc ttctttctct gtaatccttt cgttgagttt tgtttccgat
     6841 caatcatagg tagttttctt gtttcgaagc atgagatcta ggaatttttt gtgattttcc
     6901 aaaattgaga tcggtttgaa attgaatttt acagcttgaa tctcagatct tgttttatca
     6961 atgttttcga tggcttgcga tgtagatcta tgataattgt ggttcagttt tgttaggaat
     7021 cgatttcggt ttagcaattg cagattaatt agggtttcca attgaattct tcagatccgt
     7081 tatggaatta tgtcaaataa tttattcaaa ttggaaatta ttgttagatc cactcttaat
     7141 ctgtttgatc caagcttcaa ttagggtttt cacttgtttc aatttcttgt tatggattct
     7201 gatttatctg ttgatgttag atccactctt aatctgtttg atccaagcgt taattagggt
     7261 tttcacttgt ttcaatttcg tgtgttggat tctgatttat ctgttgttga tgtgattaca
     7321 gaatgaagtt catcaagacc cttttccttg tgaccttgct aatggctttg ggcgctatta
     7381 cactatcatc agcaacatct tctaaaaacg aagaaccaaa ctctctccaa ggaacaagcc
     7441 acttcctttc ccggaaacaa aacagaatct ctgtgtcatg taacaaatat ccaaagattt
     7501 gcaatatcaa aggtagtcca ggccctaatt gctgcaacaa caattgtgtc aacttttcca
     7561 tagacatgtt taattgtggg aggtgtggaa agaaatgtag ctttccaaag atatgttgtg
     7621 aaggtaaatg tgtgaatcct aggagtaacg agaagcattg tggaaaatgt ggtaacaagt
     7681 gtgacaatag aggttcatgt gtttatggaa tgtgcagata tgcgtaggct tctctagcta
     7741 gagtcgatcg acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg
     7801 aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
     7861 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
     7921 tactaaaatc cagatcgcta agcttgaatt cttgcggaga gcggagaatt aagggagtca
     7981 cgttatgacc cccgccgatg acgcgggac
//
