LOCUS       sequence-325457-        7835 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 7835)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7835)
  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            "unpublished"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:MtIDA11:T35s
FEATURES             Location/Qualifiers
     source          1..7835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      139..333
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(399..1472)
                     /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"
     CDS             complement(1901..2530)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    2794..2818
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(2913..3165)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(3178..3972)
                     /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(3289..3308)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3899..3918
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3994..4173)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     polyA_signal    4273..4447
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5886..5910
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6033..6621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6113..6132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6792..7586)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7587..7691)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgcctggcc gtaggccagc catttttgag cggccagcgg ccgcgatagg ccgacgcgaa
       61 gcggcggggc gtagggagcg cagcgaccga agggtaggcg ctttttgcag ctcttcggct
      121 gtgcgctggc cagacagtta tgcacaggcc aggcgggttt taagagtttt aataagtttt
      181 aaagagtttt aggcggaaaa atcgcctttt ttctctttta tatcagtcac ttacatgtgt
      241 gaccggttcc caatgtacgg ctttgggttc ccaatgtacg ggttccggtt cccaatgtac
      301 ggctttgggt tcccaatgta cgtgctatcc acaggaaaga agctttttcg acctttttcc
      361 cctgctaggg caatttgccc tagcatctgc tccgtacatt aggaaccggc ggatgcttcg
      421 ccctcgatca ggttgcggta gcgcatgact aggatcgggc cagcctgccc cgcctcctcc
      481 ttcaaatcgt actccggcag gtcatttgac ccgatcagct tgcgcacggt gaaacagaac
      541 ttcttgaact ctccggcgct gccactgcgt tcgtagatcg tcttgaacaa ccatctggct
      601 tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa aacggccgat gccgggatcg
      661 atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg
      721 cggtacatcc aatcagctag ctcgatctcg atgtactccg gccgcccggt ttcgctcttt
      781 acgatcttgt agcggctaat caaggcttca ccctcggata ccgtcaccag gcggccgttc
      841 ttggccttct tcgtacgctg catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct
      901 accaggtcgt ctttctgctt tccgccatcg gctcgccggc agaacttgag tacgtccgca
      961 acgtgtggac ggaacacgcg gccgggcttg tctcccttcc cttcccggta tcggttcatg
     1021 gattcggtta gatgggaaac cgccatcagt accaggtcgt aatcccacac actggccatg
     1081 ccggccggcc ctgcggaaac ctctacgtgc ccgtctggaa gctcgtagcg gatcacctcg
     1141 ccagctcgtc ggtcacgctt cgacagacgg aaaacggcca cgtccatgat gctgcgacta
     1201 tcgcgggtgc ccacgtcata gagcatcgga acgaaaaaat ctggttgctc gtcgcccttg
     1261 ggcggcttcc taatcgacga cgcaccggct gccggcggtt gccgggattc tttgcggatt
     1321 cgatcagcgg ccgcttgcca cgattcaccg gggcgtgctt ctgcctcgat gcgttgccgc
     1381 tgggcggcct gcgcggcctt caacttctcc accaggtcat cacccagcgc cgcgccgatt
     1441 tgtaccgggc cggatggttt gcgaccgctc acgccgattc ctcgggcttg ggggttccag
     1501 tgccattgca gggccggcag acaacccagc cgcttacgcc tggccaaccg cccgttcctc
     1561 cacacatggg gcattccacg gcgtcggtgc ctggttgttc ttgattttcc atgccgcctc
     1621 ctttagccgc taaaattcat ctactcattt attcatttgc tcatttactc tggtagctgc
     1681 gcgatgtatt cagatagcag ctcggtaatg gtcttgcctt ggcgtaccgc gtacatcttc
     1741 agcttggtgt gatcctccgc cggcaactga aagttgaccc gcttcatggc tggcgtgtct
     1801 gccaggctgg ccaacgttgc agccttgctg ctgcgtgcgc tcggacggcc ggcacttagc
     1861 gtgtttgtgc ttttgctcat tttctcttta cctcattaac tcaaatgagt tttgatttaa
     1921 tttcagcggc cagcgcctgg acctcgcggg cagcgtcgcc ctcgggttct gattcaagaa
     1981 cggttgtgcc ggcggcggca gtgcctgggt agctcacgcg ctgcgtgata cgggactcaa
     2041 gaatgggcag ctcgtacccg gccagcgcct cggcaacctc accgccgatg cgcgtgcctt
     2101 tgatcgcccg cgacacgaca aaggccgctt gtagccttcc atccgtgacc tcaatgcgct
     2161 gcttaaccag ctccaccagg tcggcggtgg cccatatgtc gtaagggctt ggctgcaccg
     2221 gaatcagcac gaagtcggct gccttgatcg cggacacagc caagtccgcc gcctggggcg
     2281 ctccgtcgat cactacgaag tcgcgccggc cgatggcctt cacgtcgcgg tcaatcgtcg
     2341 ggcggtcgat gccgacaacg gttagcggtt gatcttcccg cacggccgcc caatcgcggg
     2401 cactgccctg gggatcggaa tcgactaaca gaacatcggc cccggcgagt tgcagggcgc
     2461 gggctagatg ggttgcgatg gtcgtcttgc ctgacccgcc tttctggtta agtacagcga
     2521 taaccttcat gcgttcccct tgcgtatttg tttatttact catcgcatca tatacgcagc
     2581 gaccgcatga cgcaagctgt tttactcaaa tacacatcac ctttttagac ggcggcgctc
     2641 ggtttcttca gcggccaagc tggccggcca ggccgccagc ttggcatcag acaaaccggc
     2701 caggatttca tgcagccgca cggctgatgg gctgcctgta tcgagtggtg attttgtgcc
     2761 gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa
     2821 attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact ggggtggatg
     2881 cagtgggccc tgccgaattc ggatccagcg tcgatctagt aacatagatg acaccgcgcg
     2941 cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa
     3001 ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat
     3061 tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac
     3121 aggattcaat cttaagaaac tttattgcca aatgtttgaa cgatctgctt gacaagctca
     3181 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     3241 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     3301 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     3361 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccct gggtcacgac
     3421 gagatcctcg ccgtcgggca tccgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     3481 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     3541 tcctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     3601 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     3661 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     3721 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     3781 tgcctcgtct tggagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     3841 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     3901 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     3961 ttgttcaatc atttgagagt gaatatgaga ctctaattgg ataccgaggg gaatttatgg
     4021 aacgtcagtg gagcattttt gacaagaaat atttgctagc tgatagtgac cttaggcgac
     4081 ttttgaacgc gcaataatgg tttctgacgt atgtgcttag ctcattaaac tccagaaacc
     4141 cgcggctgag tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg
     4201 tcccgcgtca tcggcggggg tcataacgtg actcccttaa ttctccgctc tccgcaagaa
     4261 ttcaagctta gcgatctgga ttttagtact ggattttggt tttaggaatt agaaatttta
     4321 ttgatagaag tattttacaa atacaaatac atactaaggg tttcttatat gctcaacaca
     4381 tgagcgaaac cctataggaa ccctaattcc cttatctggg aactactcac acattattat
     4441 ggagaaactc gagcttgtcg atcgactcta gctagagaag ctcatggtga tctcaaactt
     4501 tgcaagccaa tgtcattgtg tttccttgaa ggagaagagt agggtatagg tattcttctt
     4561 ggcaagaaac caaaaaagtg acccttgtgc tcatactttg gcttcacttt gaacacatta
     4621 gtggctcttg agccatcaca gtgaccaaat atgcagagaa acaagagcaa taaaagcaat
     4681 gaagtttgaa gttttctact atgtagaacc attctgtaat cacatcaaca acagataaat
     4741 cagaatccaa cacacgaaat tgaaacaagt gaaaacccta attaacgctt ggatcaaaca
     4801 gattaagagt ggatctaaca tcaacagata aatcagaatc cataacaaga aattgaaaca
     4861 agtgaaaacc ctaattgaag cttggatcaa acagattaag agtggatcta acaataattt
     4921 ccaatttgaa taaattattt gacataattc cataacggat ctgaagaatt caattggaaa
     4981 ccctaattaa tctgcaattg ctaaaccgaa atcgattcct aacaaaactg aaccacaatt
     5041 atcatagatc tacatcgcaa gccatcgaaa acattgataa aacaagatct gagattcaag
     5101 ctgtaaaatt caatttcaaa ccgatctcaa ttttggaaaa tcacaaaaaa ttcctagatc
     5161 tcatgcttcg aaacaagaaa actacctatg attgatcgga aacaaaactc aacgaaagga
     5221 ttacagagaa agaagagaga aattggatca taccttgaaa gtagagagaa gcgatatgaa
     5281 caagaattgt tagggttggg aactgaattg tgaagaagct aaggggattg gaggtaattt
     5341 atagcaaaag cggtgaaatt attaaccagt caaattgtga cacgtgtcct tatgacgcgt
     5401 cactagttgc gtgtggtttg ggataggata cgttgtgtgg ttgagttctt cttggaaacg
     5461 gtagttgcaa attgcaagtt gttggcggtt tgttaatttc acggagttaa acgcatctcg
     5521 ccgccttttg agtttggatt gataaatcgt tacaacggcg ttactttgct gggccccgtt
     5581 ttttttgaat tgggccaccg caccagtgca agttgggggt tccttggttt tccagaatgg
     5641 gctccactct ctatgattga ttggaggagg ccggctgctc agaaatttta taataagttg
     5701 ttatatactt ctccccttaa aataagtgat ttttgtctat ccatagttgc aataagttgt
     5761 actaacaact tatctaaatt actttatcaa taatacaagt aattcaatta attatttcct
     5821 caaaatcctc tctccctcca ctagaggatg cacatgtgac cgagggagtt taaactatca
     5881 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
     5941 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgccaaccac
     6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6781 tggtctgaca gctaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca
     6841 ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc
     6901 ctgatcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca
     6961 agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg
     7021 ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg
     7081 cgcggatctt taaatggagt gtcctcttcc cagttttcgc aatccacatc ggccagatcg
     7141 ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa
     7201 tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt
     7261 tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg
     7321 agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt
     7381 ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac
     7441 cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta
     7501 gcaggagaca ttccttccgt atcttttgcg cagcggtatt tttcgatcag ttttttcaat
     7561 tccggtgata ttctcatttt agccatactc ttcctttttc aatattattg aagcatttat
     7621 caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7681 ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg
     7741 aggggcgcag cccctggggg gatgggagct cggtgccttg atgtgggcgc cggcggtcga
     7801 gtggcgacgg cgcggcttgt ccgcgccctg gtaga
//
