LOCUS       sequence-386957-        8999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 gene expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8999)
  TITLE     Patron Lab Plasmids
REFERENCE   2  (bases 1 to 8999)
  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"
COMMENT     Sequence Label: pEPOZ1KN0146
FEATURES             Location/Qualifiers
     source          1..8999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      117..311
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(547..569)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    655..795
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     710..729
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(810..827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(981..1569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1061..1080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1657..2472)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1657..2472)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2380..2399
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2905..2929
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3360..3705
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3648..3668
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    5399..5573
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5594..5618
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6919..7548
                     /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(7977..8999,1..51)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta atgtacggag
       61 cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggact ctttcctgtg
      121 gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg
      181 aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa
      241 aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg
      301 gcctgtgcat aactgtctgg ccagcgcaca gccgaagtgc tgcaaaaagc gcctaccctt
      361 cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc
      421 tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg
      481 ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg
      541 tgaaaacctc tgacacatgc agctcccggt gacggtcaca gcttgtctgt aagcggatgc
      601 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
      661 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
      721 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      781 aaataccgca tcaggcgctc atccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1381 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     1441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1621 tcacgttaag ggattttggt catgcattct aggtgattag aaaaactcat cgagcatcaa
     1681 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1741 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1801 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1861 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1921 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1981 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagtc gaaatacgcg
     2041 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2101 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2161 tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2221 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2281 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2341 atacaatcgg tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2401 atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg tttcccgttg
     2461 aatatggctc ataacagaac ttattatttc cttcctcttt tctacagtat ttaaagatac
     2521 cccaagaagc taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc
     2581 ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat
     2641 cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca
     2701 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
     2761 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
     2821 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     2881 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca taacgaattg
     2941 ctcttcagca aggaggtcaa catggtggag cacgacactc tggtctactc caaaaatgtc
     3001 aaagatacag tctcagaaga tcaaagggct attgagactt ttcaacaaag gataatttcg
     3061 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa
     3121 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat
     3181 ctctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     3241 gaagaggttc caaccacgtc tacaaagcaa gtggattgat gtgataacat ggtggagcac
     3301 gacactctgg tctactccaa aaatgtcaaa gatacagtct cagaagatca aagggctatt
     3361 gagacttttc aacaaaggat aatttcggga aacctcctcg gattccattg cccagctatc
     3421 tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg ccatcattgc
     3481 gataaaggaa aggctatcat tcaagatctc tctgccgaca gtggtcccaa agatggaccc
     3541 ccacccacga ggagcatcgt ggaaaaagaa gaggttccaa ccacgtctac aaagcaagtg
     3601 gattgatgtg acatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     3661 gacccttcct ctatataagg aagttcattt catttggaga ggacacgctc gagtataaga
     3721 gctcattttt acaacaatta ccaacaacaa caaacaacaa acaacattac aattacattt
     3781 acaattatcg atacaatggg gagagggaag atagttatac gaaggatcga taactctaca
     3841 agtagacaag tgactttctc caagagaagg agtggtttgc ttaagaaagc taaagagtta
     3901 tcgatccttt gtgatgcaga agttggtgtt atcatattct ctagcaccgg aaagctctac
     3961 gactacgcaa gcaattcaag tatgaaaaca atcattgagc ggtacaacag agtaaaagag
     4021 gagcagcatc aacttctgaa tcatgcctca gagataaagt tttggcaaag agaggttgca
     4081 agtttgcagc agcagctcca atatctacaa gaatgccaca ggaaactagt gggagaggaa
     4141 ctttctggaa tgaatgctaa cgacctacaa aaccttgaag accagctagt aacaagtcta
     4201 aaaggtgttc gtctcaaaaa ggatcaactt atgacaaatg aaatcagaga acttaatcgt
     4261 aagggacaaa tcatccaaaa agagaatcac gagctacaaa atattgtaga tataatgcgt
     4321 aaggaaaata ttaaattgca aaagaaggtt catggaagaa caaatgcgat tgaaggcaat
     4381 tcaagtgtag atccaataag caatggaacc acaacatatg caccaccgca acttcaactc
     4441 atacaactac aaccagctcc tagagaaaaa tcaatcagac taggaatatg tctgaatctt
     4501 ggttcgggcg ggagcggtgg aagcggcgaa attcagcaag ccactgcagg agtctcacaa
     4561 gacacttcgg aaaatcctaa caaaacaata gttcctgcag cattaccaca gctcacccct
     4621 accttggtgt cactgctgga ggtgattgaa cccgaggtgt tgtatgcagg atatgatagc
     4681 tctgttccag attcagcatg gagaattatg accacactca acatgttagg tgggcgtcaa
     4741 gtgattgcag cagtgaaatg ggcaaaggcg atactaggct tgagaaactt acacctcgat
     4801 gaccaaatga ccctgctaca gtactcatgg atgtttctca tggcatttgc cttgggttgg
     4861 agatcataca gacaatcaag cggaaacctg ctctgctttg ctcctgatct gattattaat
     4921 gagcagagaa tgtctctacc ctgcatgtat gaccaatgta aacacatgct gtttgtctcc
     4981 tctgaattac aaagattgca ggtatcctat gaagagtatc tctgtatgaa aaccttactg
     5041 cttctctcct cagttcctaa ggaaggtctg aagagtcaag agttatttga tgagattcga
     5101 atgacttata tcaaagagct aggaaaagcc atcgtcaaaa gggaagggaa ctccagtcag
     5161 aactggcaac ggttttacca actgacaaag cttctggact ccatgcatga ggtggttgag
     5221 aatctcctta cctactgctt ccagacattt ttggataaga ccatgagtat tgaattccca
     5281 gagatgttag ctgaaatcat cactaatcag ataccaaaat attcaaatgg aaatatcaaa
     5341 aagcttctgt ttcatcaaaa atgagcttct ctagctagag tcgatcgaca agctcgagtt
     5401 tctccataat aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc
     5461 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
     5521 atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag atcgctatac
     5581 tgaagagcaa ttctgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
     5641 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     5701 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct
     5761 atagtgcagt cggcttctga cgttcagtgc agccgtcatc tgaaaacgac atgtcgcaca
     5821 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt
     5881 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca
     5941 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga
     6001 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca
     6061 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg
     6121 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca
     6181 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg
     6241 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg
     6301 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg
     6361 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga
     6421 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga
     6481 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg
     6541 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac
     6601 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac
     6661 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacct
     6721 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
     6781 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
     6841 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
     6901 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
     6961 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
     7021 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
     7081 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
     7141 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
     7201 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
     7261 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
     7321 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
     7381 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
     7441 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
     7501 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
     7561 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
     7621 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
     7681 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
     7741 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
     7801 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
     7861 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
     7921 tgggttgcct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
     7981 gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
     8041 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
     8101 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
     8161 ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
     8221 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
     8281 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
     8341 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
     8401 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
     8461 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
     8521 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
     8581 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
     8641 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
     8701 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
     8761 tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc
     8821 tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac
     8881 gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt
     8941 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatccta
//
