LOCUS       sequence-324505-        8231 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 8231)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8231)
  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            "unpublished"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:MtFam4_1:T35s
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      313..507
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        790..894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             895..1689
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1860..2448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2349..2368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2571..2595
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    4430..4604
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        4704..4883
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4905..5699
                     /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(4959..4978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5569..5588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5712..5964
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6059..6083
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6347..6976
                     /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(7405..8231,1..247)
                     /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 ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt
       61 caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt
      121 gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca
      181 ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg
      241 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
      301 aagcttcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
      361 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
      421 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
      481 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
      541 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
      601 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
      661 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccg agctcccatc
      721 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      781 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      841 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatggct
      901 aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg cgcaaaagat
      961 acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga aaacctatat
     1021 ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg ggaaaaggac
     1081 atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt tgaacggcat
     1141 gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc ggaagagtat
     1201 gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat caggctcttt
     1261 cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg cttagccgaa
     1321 ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg ggaagaggac
     1381 actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa gcccgaagag
     1441 gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa agatggcaaa
     1501 gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta tgacattgcc
     1561 ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga gctatttttt
     1621 gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt actggatgaa
     1681 ttgttttagc tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1801 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1861 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1921 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1981 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2041 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2101 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2161 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2221 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2281 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2341 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2401 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2461 gcggcctttt tacggttcct gtggttggca catacaaatg gacgaacgga taaacctttt
     2521 cacgcccttt taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc
     2581 caatatatcc tgtcaaacac tgatagttta aactccctcg gtcacatgtg catcctctag
     2641 tggagggaga gaggattttg aggaaataat taattgaatt acttgtatta ttgataaagt
     2701 aatttagata agttgttagt acaacttatt gcaactatgg atagacaaaa atcacttatt
     2761 ttaaggggag aagtatataa caacttatta taaaatttct gagcagccgg cctcctccaa
     2821 tcaatcatag agagtggagc ccattctgga aaaccaagga acccccaact tgcactggtg
     2881 cggtggccca attcaaaaaa aacggggccc agcaaagtaa cgccgttgta acgatttatc
     2941 aatccaaact caaaaggcgg cgagatgcgt ttaactccgt gaaattaaca aaccgccaac
     3001 aacttgcaat ttgcaactac cgtttccaag aagaactcaa ccacacaacg tatcctatcc
     3061 caaaccacac gcaactagtg acgcgtcata aggacacgtg tcacaatttg actggttaat
     3121 aatttcaccg cttttgctat aaattacctc caatcccctt agcttcttca caattcagtt
     3181 cccaacccta acaattcttg ttcatatcgc ttctctctac tttcaaggta tgatccaatt
     3241 tctctcttct ttctctgtaa tcctttcgtt gagttttgtt tccgatcaat cataggtagt
     3301 tttcttgttt cgaagcatga gatctaggaa ttttttgtga ttttccaaaa ttgagatcgg
     3361 tttgaaattg aattttacag cttgaatctc agatcttgtt ttatcaatgt tttcgatggc
     3421 ttgcgatgta gatctatgat aattgtggtt cagttttgtt aggaatcgat ttcggtttag
     3481 caattgcaga ttaattaggg tttccaattg aattcttcag atccgttatg gaattatgtc
     3541 aaataattta ttcaaattgg aaattattgt tagatccact cttaatctgt ttgatccaag
     3601 cttcaattag ggttttcact tgtttcaatt tcttgttatg gattctgatt tatctgttga
     3661 tgttagatcc actcttaatc tgtttgatcc aagcgttaat tagggttttc acttgtttca
     3721 atttcgtgtg ttggattctg atttatctgt tgttgatgtg attacagaat gattcaaatc
     3781 attttcctcc tttcactttt cctatccatt tccaatgctt ctgttcaaga tttctgtgta
     3841 gcagatatca aaggctcaga cactccttca ggctaccctt gcaagccagc ttcaacagta
     3901 acctcagatg actttgcatt tgaaggctta attgcacctg gaaacataac aaacattatc
     3961 aacgccgccg ttacacctgc atttgttgct caatttccag ctgttaatgg tcttggtctc
     4021 tctgcagcaa ggttagacct tggtcctgct ggtgttatcc cacttcacac tcaccctggt
     4081 gccagtgaac ttcttgttgt gacacaaggt cacatcactg ctggatttgt ttcatcagct
     4141 aacactgttt atatcaagac attgaagaaa ggtgagctta tggtttttcc acaaggtttg
     4201 ttgcattttc aagctactgc tggtaagaga aatgctgttg cttttgctgt ttttagtagt
     4261 gctagtcctg gacttcagat tcttgatttt gcactttttg caagtaactt ctctactcct
     4321 ttgatcacta agaccacttt ccttgatcct gttcttgtca agaagcttaa gagtattctt
     4381 ggtggcagtg gttaagcttc tctagctaga gtcgatcgac aagctcgagt ttctccataa
     4441 taatgtgtga gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg
     4501 ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa
     4561 atttctaatt cctaaaacca aaatccagta ctaaaatcca gatcgctaag cttgaattct
     4621 tgcggagagc ggagaattaa gggagtcacg ttatgacccc cgccgatgac gcgggacaag
     4681 ccgttttacg tttggaactg acagaaccgc aacgttgaag gagccactca gccgcgggtt
     4741 tctggagttt aatgagctaa gcacatacgt cagaaaccat tattgcgcgt tcaaaagtcg
     4801 cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa atgctccact gacgttccat
     4861 aaattcccct cggtatccaa ttagagtctc atattcactc tcaaatgatt gaacaagatg
     4921 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4981 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5041 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actccaagac gaggcagcgc
     5101 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5161 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5221 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5281 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgaggacgta
     5341 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5401 cgccagccga actgttcgcc aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg
     5461 tgacccaggg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5521 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5581 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5641 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5701 cttgtcaagc agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     5761 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     5821 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     5881 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     5941 cggtgtcatc tatgttacta gatcgacgct ggatccgaat tcggcagggc ccactgcatc
     6001 caccccagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt
     6061 ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca
     6121 caaaatcacc actcgataca ggcagcccat cagccgtgcg gctgcatgaa atcctggccg
     6181 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
     6241 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
     6301 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
     6361 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
     6421 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
     6481 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     6541 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     6601 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     6661 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     6721 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     6781 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     6841 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     6901 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     6961 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     7021 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     7081 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     7141 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     7201 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     7261 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     7321 aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg
     7381 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
     7441 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     7501 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     7561 cgcaaagaat cccggcaacc gccggcagcc ggtgcgtcgt cgattaggaa gccgcccaag
     7621 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     7681 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     7741 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     7801 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     7861 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     7921 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     7981 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     8041 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     8101 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     8161 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     8221 cccggcatcg g
//
