LOCUS       sequence-324486-        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  JOURNAL   unpublished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            "unpublished"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:MtCEP1:T35s
FEATURES             Location/Qualifiers
     source          1..8009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        203..307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             308..1102
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1273..1861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1762..1781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1984..2008
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    3621..3795
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        3895..4074
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4096..4890
                     /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(4150..4169)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4760..4779
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4903..5155
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5250..5274
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5538..6167
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6596..7669
                     /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      7735..7929
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc
       61 taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca
      121 ccgagctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc accccaaaaa
      181 tggcagcgct ggccaattcg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      241 aaatatgtat ccgctcatga gacaataacc ttgataaatg cttcaataat attgaaaaag
      301 gaagagtatg gctaaaatga gaatatcacc ggaattgaaa aaactgatcg aaaaataccg
      361 ctgcgcaaaa gatacggaag gaatgtctcc tgctaaggta tataagctgg tgggagaaaa
      421 tgaaaaccta tatttaaaaa tgacggacag ccggtataaa gggaccacct atgatgtgga
      481 acgggaaaag gacatgatgc tatggctgga aggaaagctg cctgttccaa aggtcctgca
      541 ctttgaacgg catgatggct ggagcaatct gctcatgagt gaggccgatg gcgtcctttg
      601 ctcggaagag tatgaagatg aacaaagccc tgaaaagatt atcgagctgt atgcggagtg
      661 catcaggctc tttcactcca tcgacatatc ggattgtccc tatacgaata gcttagacag
      721 ccgcttagcc gaattggatt acttactgaa taacgatctg gccgatgtgg attgcgaaaa
      781 ctgggaagag gacactccat ttaaagatcc gcgcgagctg tatgattttt taaagacgga
      841 aaagcccgaa gaggaacttg tcttttccca cggcgacctg ggagacagca acatctttgt
      901 gaaagatggc aaagtaagtg gctttattga tcttgggaga agcggcaggg cggacaagtg
      961 gtatgacatt gccttctgcg tccggtcgat cagggaggat atcggggaag aacagtatgt
     1021 cgagctattt tttgacttac tggggatcaa gcctgattgg gagaaaataa aatattatat
     1081 tttactggat gaattgtttt agctgtcaga ccaagtttac tcatatatac tttagattga
     1141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1381 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     1441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1861 aaacgccagc aacgcggcct ttttacggtt cctgtggttg gcacatacaa atggacgaac
     1921 ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct
     1981 taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaactccc tcggtcacat
     2041 gtgcatcctc tagtggaggg agagaggatt ttgaggaaat aattaattga attacttgta
     2101 ttattgataa agtaatttag ataagttgtt agtacaactt attgcaacta tggatagaca
     2161 aaaatcactt attttaaggg gagaagtata taacaactta ttataaaatt tctgagcagc
     2221 cggcctcctc caatcaatca tagagagtgg agcccattct ggaaaaccaa ggaaccccca
     2281 acttgcactg gtgcggtggc ccaattcaaa aaaaacgggg cccagcaaag taacgccgtt
     2341 gtaacgattt atcaatccaa actcaaaagg cggcgagatg cgtttaactc cgtgaaatta
     2401 acaaaccgcc aacaacttgc aatttgcaac taccgtttcc aagaagaact caaccacaca
     2461 acgtatccta tcccaaacca cacgcaacta gtgacgcgtc ataaggacac gtgtcacaat
     2521 ttgactggtt aataatttca ccgcttttgc tataaattac ctccaatccc cttagcttct
     2581 tcacaattca gttcccaacc ctaacaattc ttgttcatat cgcttctctc tactttcaag
     2641 gtatgatcca atttctctct tctttctctg taatcctttc gttgagtttt gtttccgatc
     2701 aatcataggt agttttcttg tttcgaagca tgagatctag gaattttttg tgattttcca
     2761 aaattgagat cggtttgaaa ttgaatttta cagcttgaat ctcagatctt gttttatcaa
     2821 tgttttcgat ggcttgcgat gtagatctat gataattgtg gttcagtttt gttaggaatc
     2881 gatttcggtt tagcaattgc agattaatta gggtttccaa ttgaattctt cagatccgtt
     2941 atggaattat gtcaaataat ttattcaaat tggaaattat tgttagatcc actcttaatc
     3001 tgtttgatcc aagcttcaat tagggttttc acttgtttca atttcttgtt atggattctg
     3061 atttatctgt tgatgttaga tccactctta atctgtttga tccaagcgtt aattagggtt
     3121 ttcacttgtt tcaatttcgt gtgttggatt ctgatttatc tgttgttgat gtgattacag
     3181 aatgaaatat ttcgctcttt ttcttgccct aattgcatgc aactattccc ttcaatctca
     3241 tgcaaggctc attaaaccat cgaaccatca caatgttcca atttcaacat cagagaagaa
     3301 agttgagtca acaataaaat caaacaatga agtagctagt tattttggag attcaagtga
     3361 agctcacacg aatgcattcc aaccaacaac accaggaaat agtcctggtg ttggtcatag
     3421 atattttacc gatgaagata tcgacgtgaa ttcgaaaaag acggtagctc agagcaaaga
     3481 tgataataaa tatgtgactg aggatactac aaatgagttc caaaaaacaa accctggtca
     3541 cagtcctggt gttggtcatt cttaccaaaa caaaattgga aattgagctt ctctagctag
     3601 agtcgatcga caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga
     3661 attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
     3721 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
     3781 actaaaatcc agatcgctaa gcttgaattc ttgcggagag cggagaatta agggagtcac
     3841 gttatgaccc ccgccgatga cgcgggacaa gccgttttac gtttggaact gacagaaccg
     3901 caacgttgaa ggagccactc agccgcgggt ttctggagtt taatgagcta agcacatacg
     3961 tcagaaacca ttattgcgcg ttcaaaagtc gcctaaggtc actatcagct agcaaatatt
     4021 tcttgtcaaa aatgctccac tgacgttcca taaattcccc tcggtatcca attagagtct
     4081 catattcact ctcaaatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4141 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4201 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4261 gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4321 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4381 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4441 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4501 caagcgaaac atcgcatcga gcgaggacgt actcggatgg aagccggtct tgtcgatcag
     4561 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4621 gcgcggatgc ccgacggcga ggatctcgtc gtgacccagg gcgatgcctg cttgccgaat
     4681 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4741 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4801 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4861 ttctatcgcc ttcttgacga gttcttctga gcttgtcaag cagatcgttc aaacatttgg
     4921 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt
     4981 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     5041 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     5101 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgacgc
     5161 tggatccgaa ttcggcaggg cccactgcat ccaccccagt acattaaaaa cgtccgcaat
     5221 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
     5281 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
     5341 tcagccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg
     5401 ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt
     5461 gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa
     5521 atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca
     5581 agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt
     5641 tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac
     5701 cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc
     5761 ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga
     5821 tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca
     5881 ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag
     5941 cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg
     6001 cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc
     6061 caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc
     6121 gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa
     6181 agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag
     6241 caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca
     6301 gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat
     6361 taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa
     6421 tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca
     6481 ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct
     6541 gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag
     6601 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga
     6661 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg
     6721 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc
     6781 cggtgcgtcg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc
     6841 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg
     6901 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca
     6961 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact
     7021 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa
     7081 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga
     7141 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt
     7201 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga
     7261 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga
     7321 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct
     7381 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct
     7441 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg
     7501 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc
     7561 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt
     7621 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca
     7681 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaagcttct ttcctgtgga
     7741 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa
     7801 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa
     7861 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc
     7921 ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg
     7981 gtcgctgcgc tccctacgcc ccgccgctt
//
