LOCUS       sequence-325450-        7802 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 7802)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7802)
  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            "unpublished"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:MtMEG2:T35s
FEATURES             Location/Qualifiers
     source          1..7802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        210..314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             315..1109
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1280..1868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1769..1788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1991..2015
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    3421..3595
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        3695..3874
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             3896..4690
                     /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(3950..3969)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4560..4579
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4703..4955
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5050..5074
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5338..5967
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6396..7469
                     /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      7535..7729
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca
       61 ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat
      121 caaggcaccg agctcccatc cccccagggg ctgcgcccct cggccgcgaa cggcctcacc
      181 ccaaaaatgg cagcgctggc caattcgtgc gcggaacccc tatttgttta tttttctaaa
      241 tacattcaaa tatgtatccg ctcatgagac aataaccttg ataaatgctt caataatatt
      301 gaaaaaggaa gagtatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa
      361 aataccgctg cgcaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg
      421 gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg
      481 atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg
      541 tcctgcactt tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg
      601 tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg
      661 cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct
      721 tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt
      781 gcgaaaactg ggaagaggac actccattta aagatccgcg cgagctgtat gattttttaa
      841 agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca
      901 tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg
      961 acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac
     1021 agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat
     1081 attatatttt actggatgaa ttgttttagc tgtcagacca agtttactca tatatacttt
     1141 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1201 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1261 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1321 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1381 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1801 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1861 tatggaaaaa cgccagcaac gcggcctttt tacggttcct gtggttggca catacaaatg
     1921 gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa taaacgctct
     1981 tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta aactccctcg
     2041 gtcacatgtg catcctctag tggagggaga gaggattttg aggaaataat taattgaatt
     2101 acttgtatta ttgataaagt aatttagata agttgttagt acaacttatt gcaactatgg
     2161 atagacaaaa atcacttatt ttaaggggag aagtatataa caacttatta taaaatttct
     2221 gagcagccgg cctcctccaa tcaatcatag agagtggagc ccattctgga aaaccaagga
     2281 acccccaact tgcactggtg cggtggccca attcaaaaaa aacggggccc agcaaagtaa
     2341 cgccgttgta acgatttatc aatccaaact caaaaggcgg cgagatgcgt ttaactccgt
     2401 gaaattaaca aaccgccaac aacttgcaat ttgcaactac cgtttccaag aagaactcaa
     2461 ccacacaacg tatcctatcc caaaccacac gcaactagtg acgcgtcata aggacacgtg
     2521 tcacaatttg actggttaat aatttcaccg cttttgctat aaattacctc caatcccctt
     2581 agcttcttca caattcagtt cccaacccta acaattcttg ttcatatcgc ttctctctac
     2641 tttcaaggta tgatccaatt tctctcttct ttctctgtaa tcctttcgtt gagttttgtt
     2701 tccgatcaat cataggtagt tttcttgttt cgaagcatga gatctaggaa ttttttgtga
     2761 ttttccaaaa ttgagatcgg tttgaaattg aattttacag cttgaatctc agatcttgtt
     2821 ttatcaatgt tttcgatggc ttgcgatgta gatctatgat aattgtggtt cagttttgtt
     2881 aggaatcgat ttcggtttag caattgcaga ttaattaggg tttccaattg aattcttcag
     2941 atccgttatg gaattatgtc aaataattta ttcaaattgg aaattattgt tagatccact
     3001 cttaatctgt ttgatccaag cttcaattag ggttttcact tgtttcaatt tcttgttatg
     3061 gattctgatt tatctgttga tgttagatcc actcttaatc tgtttgatcc aagcgttaat
     3121 tagggttttc acttgtttca atttcgtgtg ttggattctg atttatctgt tgttgatgtg
     3181 attacagaat gatgatgatg aatgaaggca ggaggaaatt gggtgctttc cagatatgtg
     3241 caatgtgcac ttgctgtggt ggtgccaaag ggttgtgctt ggcctctcct tgttgctatg
     3301 tcttcacatg taaccttcca aataagcctt ttggcttttg ttccttcatc cctaagacct
     3361 gtaattgctt tggttgccat cattaggctt ctctagctag agtcgatcga caagctcgag
     3421 tttctccata ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt
     3481 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
     3541 ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcgctaa
     3601 gcttgaattc ttgcggagag cggagaatta agggagtcac gttatgaccc ccgccgatga
     3661 cgcgggacaa gccgttttac gtttggaact gacagaaccg caacgttgaa ggagccactc
     3721 agccgcgggt ttctggagtt taatgagcta agcacatacg tcagaaacca ttattgcgcg
     3781 ttcaaaagtc gcctaaggtc actatcagct agcaaatatt tcttgtcaaa aatgctccac
     3841 tgacgttcca taaattcccc tcggtatcca attagagtct catattcact ctcaaatgat
     3901 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3961 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4021 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactccaaga
     4081 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4141 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4201 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4261 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4321 gcgaggacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4381 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcggatgc ccgacggcga
     4441 ggatctcgtc gtgacccagg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4501 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4561 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4621 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4681 gttcttctga gcttgtcaag cagatcgttc aaacatttgg caataaagtt tcttaagatt
     4741 gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca
     4801 tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt
     4861 cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa
     4921 attatcgcgc gcggtgtcat ctatgttact agatcgacgc tggatccgaa ttcggcaggg
     4981 cccactgcat ccaccccagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag
     5041 cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg
     5101 gcagctcggc acaaaatcac cactcgatac aggcagccca tcagccgtgc ggctgcatga
     5161 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     5221 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     5281 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
     5341 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
     5401 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     5461 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     5521 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     5581 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     5641 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     5701 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     5761 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     5821 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     5881 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     5941 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     6001 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     6061 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     6121 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     6181 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     6241 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     6301 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgtctg ccggccctgc
     6361 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
     6421 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     6481 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     6541 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgtcg tcgattagga
     6601 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     6661 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     6721 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     6781 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     6841 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     6901 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
     6961 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
     7021 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
     7081 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
     7141 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
     7201 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
     7261 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
     7321 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
     7381 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
     7441 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
     7501 tagcagggga aaaaggtcga aaaagcttct ttcctgtgga tagcacgtac attgggaacc
     7561 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
     7621 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
     7681 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
     7741 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
     7801 cc
//
