LOCUS       sequence-324494-        7913 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 7913)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7913)
  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            "unpublished"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:MtCEP8:T35s
FEATURES             Location/Qualifiers
     source          1..7913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      928..1122
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1405..1509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1510..2304
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2475..3063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2964..2983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3186..3210
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4453..4464
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4727..4901
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5001..5180
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5202..5996
                     /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(5256..5275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5866..5885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6009..6261
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6356..6380
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6644..7273
                     /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(7702..7913,1..862)
                     /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 caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga
       61 tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg
      121 cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg
      181 catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc
      241 catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt
      301 tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt
      361 agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa
      421 gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt
      481 aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg
      541 cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat
      601 cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga
      661 agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa
      721 gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa
      781 ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg
      841 cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg
      901 ggaaaaaggt cgaaaaagct tctttcctgt ggatagcacg tacattggga acccaaagcc
      961 gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc
     1021 acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt
     1081 aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac
     1141 agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc
     1201 ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa
     1261 tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg
     1321 caccgagctc ccatcccccc aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa
     1381 aatggcagcg ctggccaatt cgtgcgcgga acccctattt gtttattttt ctaaatacat
     1441 tcaaatatgt atccgctcat gagacaataa ccttgataaa tgcttcaata atattgaaaa
     1501 aggaagagta tggctaaaat gagaatatca ccggaattga aaaaactgat cgaaaaatac
     1561 cgctgcgcaa aagatacgga aggaatgtct cctgctaagg tatataagct ggtgggagaa
     1621 aatgaaaacc tatatttaaa aatgacggac agccggtata aagggaccac ctatgatgtg
     1681 gaacgggaaa aggacatgat gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg
     1741 cactttgaac ggcatgatgg ctggagcaat ctgctcatga gtgaggccga tggcgtcctt
     1801 tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag
     1861 tgcatcaggc tctttcactc catcgacata tcggattgtc cctatacgaa tagcttagac
     1921 agccgcttag ccgaattgga ttacttactg aataacgatc tggccgatgt ggattgcgaa
     1981 aactgggaag aggacactcc atttaaagat ccgcgcgagc tgtatgattt tttaaagacg
     2041 gaaaagcccg aagaggaact tgtcttttcc cacggcgacc tgggagacag caacatcttt
     2101 gtgaaagatg gcaaagtaag tggctttatt gatcttggga gaagcggcag ggcggacaag
     2161 tggtatgaca ttgccttctg cgtccggtcg atcagggagg atatcgggga agaacagtat
     2221 gtcgagctat tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat
     2281 attttactgg atgaattgtt ttagctgtca gaccaagttt actcatatat actttagatt
     2341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2581 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3061 aaaaacgcca gcaacgcggc ctttttacgg ttcctgtggt tggcacatac aaatggacga
     3121 acggataaac cttttcacgc ccttttaaat atccgattat tctaataaac gctcttttct
     3181 cttaggttta cccgccaata tatcctgtca aacactgata gtttaaactc cctcggtcac
     3241 atgtgcatcc tctagtggag ggagagagga ttttgaggaa ataattaatt gaattacttg
     3301 tattattgat aaagtaattt agataagttg ttagtacaac ttattgcaac tatggataga
     3361 caaaaatcac ttattttaag gggagaagta tataacaact tattataaaa tttctgagca
     3421 gccggcctcc tccaatcaat catagagagt ggagcccatt ctggaaaacc aaggaacccc
     3481 caacttgcac tggtgcggtg gcccaattca aaaaaaacgg ggcccagcaa agtaacgccg
     3541 ttgtaacgat ttatcaatcc aaactcaaaa ggcggcgaga tgcgtttaac tccgtgaaat
     3601 taacaaaccg ccaacaactt gcaatttgca actaccgttt ccaagaagaa ctcaaccaca
     3661 caacgtatcc tatcccaaac cacacgcaac tagtgacgcg tcataaggac acgtgtcaca
     3721 atttgactgg ttaataattt caccgctttt gctataaatt acctccaatc cccttagctt
     3781 cttcacaatt cagttcccaa ccctaacaat tcttgttcat atcgcttctc tctactttca
     3841 aggtatgatc caatttctct cttctttctc tgtaatcctt tcgttgagtt ttgtttccga
     3901 tcaatcatag gtagttttct tgtttcgaag catgagatct aggaattttt tgtgattttc
     3961 caaaattgag atcggtttga aattgaattt tacagcttga atctcagatc ttgttttatc
     4021 aatgttttcg atggcttgcg atgtagatct atgataattg tggttcagtt ttgttaggaa
     4081 tcgatttcgg tttagcaatt gcagattaat tagggtttcc aattgaattc ttcagatccg
     4141 ttatggaatt atgtcaaata atttattcaa attggaaatt attgttagat ccactcttaa
     4201 tctgtttgat ccaagcttca attagggttt tcacttgttt caatttcttg ttatggattc
     4261 tgatttatct gttgatgtta gatccactct taatctgttt gatccaagcg ttaattaggg
     4321 ttttcacttg tttcaatttc gtgtgttgga ttctgattta tctgttgttg atgtgattac
     4381 agaatggcac aaaacaagac catagttttc tctgttattt ccctagcatt gatcattttc
     4441 tgcatgcagt cgatcgaggg gcgccttgta aaatacatcg atgaaagtaa cctcctgaag
     4501 aatgttaaac atgatggaat ttcagatgca aatgaagcta ctcttgttaa cgtgactcca
     4561 acaatattgc caccaagtgc tgtggtaggt tcaaatgggg ttgcagcacc tcctccaagt
     4621 catgatgtgg gtgcttttag acccacaacc cctgggaaca gtcctggtgt aggtcactct
     4681 attcactact aggcttctct agctagagtc gatcgacaag ctcgagtttc tccataataa
     4741 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
     4801 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
     4861 tctaattcct aaaaccaaaa tccagtacta aaatccagat cgctaagctt gaattcttgc
     4921 ggagagcgga gaattaaggg agtcacgtta tgacccccgc cgatgacgcg ggacaagccg
     4981 ttttacgttt ggaactgaca gaaccgcaac gttgaaggag ccactcagcc gcgggtttct
     5041 ggagtttaat gagctaagca catacgtcag aaaccattat tgcgcgttca aaagtcgcct
     5101 aaggtcacta tcagctagca aatatttctt gtcaaaaatg ctccactgac gttccataaa
     5161 ttcccctcgg tatccaatta gagtctcata ttcactctca aatgattgaa caagatggat
     5221 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5281 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5341 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact ccaagacgag gcagcgcggc
     5401 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5461 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5521 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5581 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga ggacgtactc
     5641 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5701 cagccgaact gttcgccagg ctcaaggcgc ggatgcccga cggcgaggat ctcgtcgtga
     5761 cccagggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5821 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5881 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5941 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagctt
     6001 gtcaagcaga tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg
     6061 gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca
     6121 tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg caattataca
     6181 tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg
     6241 tgtcatctat gttactagat cgacgctgga tccgaattcg gcagggccca ctgcatccac
     6301 cccagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta
     6361 caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa
     6421 aatcaccact cgatacaggc agcccatcag ccgtgcggct gcatgaaatc ctggccggtt
     6481 tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc
     6541 gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt
     6601 atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt
     6661 acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct
     6721 gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga
     6781 ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat
     6841 tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca
     6901 ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca
     6961 gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat
     7021 tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac
     7081 gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc
     7141 ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga
     7201 atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc
     7261 aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt
     7321 gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg
     7381 ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg
     7441 tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta
     7501 ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg
     7561 catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac
     7621 gggcggttgg ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg gcactggaac
     7681 ccccaagccc gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc
     7741 gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa
     7801 cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc
     7861 aaagaatccc ggcaaccgcc ggcagccggt gcgtcgtcga ttaggaagcc gcc
//
