LOCUS       sequence-325437-        7737 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 7737)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7737)
  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            "unpublished"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:MtIDA35:T35s
FEATURES             Location/Qualifiers
     source          1..7737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      42..630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     531..550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    753..777
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        2392..2571
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2593..3387
                     /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(2647..2666)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3257..3276
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3400..3652
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3747..3771
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4035..4664
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5093..6166
                     /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      6232..6426
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        6709..6813
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6814..7608
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
       61 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      181 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      661 ctgtggttgg cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc
      721 cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac
      781 actgatagtt taaactccct cggtcacatg tgcatcctct agtggaggga gagaggattt
      841 tgaggaaata attaattgaa ttacttgtat tattgataaa gtaatttaga taagttgtta
      901 gtacaactta ttgcaactat ggatagacaa aaatcactta ttttaagggg agaagtatat
      961 aacaacttat tataaaattt ctgagcagcc ggcctcctcc aatcaatcat agagagtgga
     1021 gcccattctg gaaaaccaag gaacccccaa cttgcactgg tgcggtggcc caattcaaaa
     1081 aaaacggggc ccagcaaagt aacgccgttg taacgattta tcaatccaaa ctcaaaaggc
     1141 ggcgagatgc gtttaactcc gtgaaattaa caaaccgcca acaacttgca atttgcaact
     1201 accgtttcca agaagaactc aaccacacaa cgtatcctat cccaaaccac acgcaactag
     1261 tgacgcgtca taaggacacg tgtcacaatt tgactggtta ataatttcac cgcttttgct
     1321 ataaattacc tccaatcccc ttagcttctt cacaattcag ttcccaaccc taacaattct
     1381 tgttcatatc gcttctctct actttcaagg tatgatccaa tttctctctt ctttctctgt
     1441 aatcctttcg ttgagttttg tttccgatca atcataggta gttttcttgt ttcgaagcat
     1501 gagatctagg aattttttgt gattttccaa aattgagatc ggtttgaaat tgaattttac
     1561 agcttgaatc tcagatcttg ttttatcaat gttttcgatg gcttgcgatg tagatctatg
     1621 ataattgtgg ttcagttttg ttaggaatcg atttcggttt agcaattgca gattaattag
     1681 ggtttccaat tgaattcttc agatccgtta tggaattatg tcaaataatt tattcaaatt
     1741 ggaaattatt gttagatcca ctcttaatct gtttgatcca agcttcaatt agggttttca
     1801 cttgtttcaa tttcttgtta tggattctga tttatctgtt gatgttagat ccactcttaa
     1861 tctgtttgat ccaagcgtta attagggttt tcacttgttt caatttcgtg tgttggattc
     1921 tgatttatct gttgttgatg tgattacaga atgggtagca ataacatgat aaagttgttc
     1981 ttgttgacaa cattaattct tcttgtattg agtatagaga acacaagtgc agcaagaaca
     2041 ctactacaaa gaggtccagt gaaaggttca acgaagaatc catgctctac agttcctggt
     2101 cgcagtaaag ggcgttgtac tttggctgaa atccatgttg ctggacatat taacgttgtt
     2161 cattcatcac caccaccttt tcctgcagat ataattccca agtttggagc agctgttatt
     2221 aataatgaac aaaaaagaga gctcatcaag accttcactg aatcagacaa acatctacaa
     2281 tttaaaaaat gagctaagct tgaattcttg cggagagcgg agaattaagg gagtcacgtt
     2341 atgacccccg ccgatgacgc gggacaagcc gttttacgtt tggaactgac agaaccgcaa
     2401 cgttgaagga gccactcagc cgcgggtttc tggagtttaa tgagctaagc acatacgtca
     2461 gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact atcagctagc aaatatttct
     2521 tgtcaaaaat gctccactga cgttccataa attcccctcg gtatccaatt agagtctcat
     2581 attcactctc aaatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2641 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2701 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2761 ctgaatgaac tccaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2821 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     2881 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     2941 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3001 gcgaaacatc gcatcgagcg aggacgtact cggatggaag ccggtcttgt cgatcaggat
     3061 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3121 cggatgcccg acggcgagga tctcgtcgtg acccagggcg atgcctgctt gccgaatatc
     3181 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3241 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3301 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3361 tatcgccttc ttgacgagtt cttctgagct tgtcaagcag atcgttcaaa catttggcaa
     3421 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
     3481 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     3541 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     3601 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgacgctgg
     3661 atccgaattc ggcagggccc actgcatcca ccccagtaca ttaaaaacgt ccgcaatgtg
     3721 ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc
     3781 caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca
     3841 gccgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg
     3901 gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag
     3961 taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata
     4021 cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga
     4081 cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag
     4141 tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc
     4201 taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc
     4261 gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca
     4321 aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg
     4381 ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg
     4441 cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc
     4501 tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag
     4561 gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg
     4621 aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga
     4681 gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa
     4741 ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt
     4801 gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac
     4861 cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga
     4921 gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg
     4981 acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg
     5041 ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg
     5101 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
     5161 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
     5221 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
     5281 tgcgtcgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
     5341 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
     5401 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
     5461 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
     5521 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
     5581 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
     5641 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
     5701 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
     5761 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
     5821 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
     5881 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
     5941 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
     6001 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
     6061 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat
     6121 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat
     6181 gctagggcaa attgccctag caggggaaaa aggtcgaaaa agcttctttc ctgtggatag
     6241 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc
     6301 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg
     6361 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg
     6421 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc
     6481 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa
     6541 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact
     6601 cgaccgccgg cgcccacatc aaggcaccga gctcccatcc ccccaggggc tgcgcccctc
     6661 ggccgcgaac ggcctcaccc caaaaatggc agcgctggcc aattcgtgcg cggaacccct
     6721 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccttga
     6781 taaatgcttc aataatattg aaaaaggaag agtatggcta aaatgagaat atcaccggaa
     6841 ttgaaaaaac tgatcgaaaa ataccgctgc gcaaaagata cggaaggaat gtctcctgct
     6901 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
     6961 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
     7021 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
     7081 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa
     7141 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
     7201 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
     7261 gatctggccg atgtggattg cgaaaactgg gaagaggaca ctccatttaa agatccgcgc
     7321 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
     7381 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
     7441 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
     7501 gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
     7561 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagct gtcagaccaa
     7621 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7681 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttc
//
