LOCUS       sequence-386960-        7946 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 7946)
  TITLE     Patron Lab Plasmids
REFERENCE   2  (bases 1 to 7946)
  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: pEPOZ1KN0222
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      310..504
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(740..762)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    848..988
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     903..922
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1003..1020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1174..1762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1254..1273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1850..2665)
                     /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(1850..2665)
                     /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     2573..2592
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3098..3122
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3553..3898
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3841..3861
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    4539..4713
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4734..4758
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6059..6688
                     /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(7117..7946,1..244)
                     /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 ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa
       61 gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg
      121 caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc
      181 tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc
      241 ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg
      301 actctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa
      361 cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga
      421 tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct
      481 taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag tgctgcaaaa
      541 agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc
      601 ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca
      661 agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt
      721 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggtgacggtc acagcttgtc
      781 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      841 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
      901 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
      961 atgcgtaagg agaaaatacc gcatcaggcg ctcatccgct tcctcgctca ctgactcgct
     1021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     1621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1801 gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtgat tagaaaaact
     1861 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1921 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1981 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2041 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2101 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2161 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2221 gtcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2281 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2341 cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2401 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2461 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2521 catcgggctt cccatacaat cggtagattg tcgcacctga ttgcccgaca ttatcgcgag
     2581 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc cttgagcaag
     2641 acgtttcccg ttgaatatgg ctcataacag aacttattat ttccttcctc ttttctacag
     2701 tatttaaaga taccccaaga agctaattat aacaagacga actccaattc actgttcctt
     2761 gcattctaaa accttaaata ccagaaaaca gctttttcaa agttgttttc aaagttggcg
     2821 tataacatag tatcgacgga gccgattttg aaaccgcggt gatcacaggc agcaacgctc
     2881 tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca
     2941 gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa
     3001 cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg
     3061 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa
     3121 acataacgaa ttgctcttca gcaaggaggt caacatggtg gagcacgaca ctctggtcta
     3181 ctccaaaaat gtcaaagata cagtctcaga agatcaaagg gctattgaga cttttcaaca
     3241 aaggataatt tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga
     3301 aaggacagta gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc
     3361 tatcattcaa gatctctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
     3421 catcgtggaa aaagaagagg ttccaaccac gtctacaaag caagtggatt gatgtgataa
     3481 catggtggag cacgacactc tggtctactc caaaaatgtc aaagatacag tctcagaaga
     3541 tcaaagggct attgagactt ttcaacaaag gataatttcg ggaaacctcc tcggattcca
     3601 ttgcccagct atctgtcact tcatcgaaag gacagtagaa aaggaaggtg gctcctacaa
     3661 atgccatcat tgcgataaag gaaaggctat cattcaagat ctctctgccg acagtggtcc
     3721 caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagaggttc caaccacgtc
     3781 tacaaagcaa gtggattgat gtgacatctc cactgacgta agggatgacg cacaatccca
     3841 ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg
     3901 ctcgagtata agagctcatt tttacaacaa ttaccaacaa caacaaacaa caaacaacat
     3961 tacaattaca tttacaatta tcgatacaat gaatagtttc gagccacaaa gccaagactc
     4021 tttgcaaaga aggtttcatc aagacaacag caccacacaa caacctcgtg acaccacgac
     4081 acctttcata cccaaaccgg cttccaaaaa ccataataat agcaactcca gctctggagc
     4141 ggccgggaga tcattccaag gctttgggct taacgtagag gacgatcttg tttcatcggt
     4201 ggttcctccg gttacggttg tgctagaggg acgttctata tgtcaacgca taagcctaga
     4261 caagcatggg agttatcaga gcttggcttc ggctctaagg caaatgtttg tcgatggagc
     4321 tgattcaacg gacgatcttg atctgtcaaa cgccattcct ggccatctta ttgcttatga
     4381 agacatggag aacgatctcc ttcttgccgg agatcttact tggaaggact ttgttcgtgt
     4441 agcgaagaga attcgaatct tgccggtcaa gggaaacaca agacaagtta aaagaaacga
     4501 gtgagcttct ctagctagag tcgatcgaca agctcgagtt tctccataat aatgtgtgag
     4561 tagttcccag ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata
     4621 agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc
     4681 ctaaaaccaa aatccagtac taaaatccag atcgctatac tgaagagcaa ttctgacagg
     4741 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
     4801 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     4861 cctcgggatc aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga
     4921 cgttcagtgc agccgtcatc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag
     4981 gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag
     5041 aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg
     5101 ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg
     5161 cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc
     5221 ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg
     5281 ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag
     5341 gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc
     5401 cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac
     5461 cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct
     5521 accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc
     5581 gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag
     5641 cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca
     5701 ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga
     5761 aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga
     5821 tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacct ctcaaccgtg cggctgcatg
     5881 aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg
     5941 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc
     6001 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat
     6061 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca
     6121 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca
     6181 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat
     6241 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat
     6301 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt
     6361 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct
     6421 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg
     6481 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct
     6541 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg
     6601 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc
     6661 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc
     6721 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc
     6781 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac
     6841 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa
     6901 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc
     6961 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
     7021 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgcct gccggccctg
     7081 caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc
     7141 ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag
     7201 gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc
     7261 gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg
     7321 aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc
     7381 acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga
     7441 cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg
     7501 ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta
     7561 accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt
     7621 ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac
     7681 gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag
     7741 aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac
     7801 aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct agctgattgg
     7861 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac
     7921 tttttgatcg atcccggcat cggccg
//
