LOCUS       sequence-386982-        9053 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 9053)
  AUTHORS   Cai YM, Witham S, Patron NJ
  TITLE     Tuning Plant Promoters Using a Simple Split Luciferase Method to 
            Assess Transcription Factor-DNA Interactions.
  JOURNAL   ACS Synth Biol. 2023 Nov 17;12(11):3482-3486. doi: 
            10.1021/acssynbio.3c00094. Epub 2023 Oct 19.
  PUBMED    37856867
REFERENCE   2  (bases 1 to 9053)
  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"; doi: 
            "10.1021/acssynbio.3c00094"; journalName: "ACS Synth Biol"; date: 
            "2023-11-17- 17"; volume: "12"; issue: "11"; pages: "3482-3486"
COMMENT     Sequence Label: pEPOZ1KN0145
FEATURES             Location/Qualifiers
     source          1..9053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(427..449)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    535..675
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     590..609
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(690..707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(861..1449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(941..960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1537..2352)
                     /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(1537..2352)
                     /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     2260..2279
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2785..2809
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3240..3585
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3528..3548
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    5333..5507
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5528..5552
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6853..7482
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7911..8984
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      join(9050..9053,1..191)
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg
       61 aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa
      121 aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg
      181 gcctgtgcat aactgtctgg ccagcgcaca gccgaagtgc tgcaaaaagc gcctaccctt
      241 cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc
      301 tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg
      361 ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg
      421 tgaaaacctc tgacacatgc agctcccggt gacggtcaca gcttgtctgt aagcggatgc
      481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
      541 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
      601 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      661 aaataccgca tcaggcgctc atccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      721 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      781 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      841 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      901 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      961 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1021 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1081 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1141 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1201 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1261 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     1321 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1381 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1441 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1501 tcacgttaag ggattttggt catgcattct aggtgattag aaaaactcat cgagcatcaa
     1561 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1621 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1681 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1741 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1801 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1861 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagtc gaaatacgcg
     1921 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     1981 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2041 tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2101 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2161 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2221 atacaatcgg tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2281 atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg tttcccgttg
     2341 aatatggctc ataacagaac ttattatttc cttcctcttt tctacagtat ttaaagatac
     2401 cccaagaagc taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc
     2461 ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat
     2521 cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca
     2581 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
     2641 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
     2701 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     2761 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca taacgaattg
     2821 ctcttcagca aggaggtcaa catggtggag cacgacactc tggtctactc caaaaatgtc
     2881 aaagatacag tctcagaaga tcaaagggct attgagactt ttcaacaaag gataatttcg
     2941 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa
     3001 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat
     3061 ctctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     3121 gaagaggttc caaccacgtc tacaaagcaa gtggattgat gtgataacat ggtggagcac
     3181 gacactctgg tctactccaa aaatgtcaaa gatacagtct cagaagatca aagggctatt
     3241 gagacttttc aacaaaggat aatttcggga aacctcctcg gattccattg cccagctatc
     3301 tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg ccatcattgc
     3361 gataaaggaa aggctatcat tcaagatctc tctgccgaca gtggtcccaa agatggaccc
     3421 ccacccacga ggagcatcgt ggaaaaagaa gaggttccaa ccacgtctac aaagcaagtg
     3481 gattgatgtg acatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     3541 gacccttcct ctatataagg aagttcattt catttggaga ggacacgctc gagtataaga
     3601 gctcattttt acaacaatta ccaacaacaa caaacaacaa acaacattac aattacattt
     3661 acaattatcg atacaatgat gaaatctggg gctgatttgc aatttccacc aggatttaga
     3721 tttcatccta cggatgagga gctagtcctc atgtatctct gtcgtaaatg cgcgtcgcag
     3781 ccgatccctg ctccgattat caccgaactc gatttgtacc gatatgatcc ttgggacctt
     3841 cccgacatgg ctttgtacgg tgaaaaggag tggtattttt tctcaccaag agatcgaaag
     3901 tatccaaacg gttcaagacc caaccgtgca gctggtactg gatattggaa agctaccgga
     3961 gctgataaac caataggtcg tcctaaaccg gttggtatta agaaggctct agtgttttac
     4021 tcgggaaaac ctccaaatgg agagaaaacc aattggatta tgcacgaata ccggctcgct
     4081 gacgttgacc ggtcggttcg taagaaaaac agtctaagat tggacgattg ggtattgtgt
     4141 cgtatatata acaagaaagg tgtcatcgag aagcgacgaa gcgatatcga ggacgggtta
     4201 aagcctgtga ctgacacgtg tccaccggaa tctgtggcga gattgatctc cggctcggag
     4261 caagcggtgt caccggaatt cacgtgtagc aacggtcggt tgagtaatgc ccttgatttt
     4321 ccgtttaatt acgtagatgc catcgccgat aacgagattg tgtcacggct attgggcggg
     4381 aatcagatgt ggtcgacgac gcttgatcca cttgtggtta ggcagggaac tttcggttcg
     4441 ggcgggagcg gtggaagcgg cgaaattcag caagccactg caggagtctc acaagacact
     4501 tcggaaaatc ctaacaaaac aatagttcct gcagcattac cacagctcac ccctaccttg
     4561 gtgtcactgc tggaggtgat tgaacccgag gtgttgtatg caggatatga tagctctgtt
     4621 ccagattcag catggagaat tatgaccaca ctcaacatgt taggtgggcg tcaagtgatt
     4681 gcagcagtga aatgggcaaa ggcgatacta ggcttgagaa acttacacct cgatgaccaa
     4741 atgaccctgc tacagtactc atggatgttt ctcatggcat ttgccttggg ttggagatca
     4801 tacagacaat caagcggaaa cctgctctgc tttgctcctg atctgattat taatgagcag
     4861 agaatgtctc taccctgcat gtatgaccaa tgtaaacaca tgctgtttgt ctcctctgaa
     4921 ttacaaagat tgcaggtatc ctatgaagag tatctctgta tgaaaacctt actgcttctc
     4981 tcctcagttc ctaaggaagg tctgaagagt caagagttat ttgatgagat tcgaatgact
     5041 tatatcaaag agctaggaaa agccatcgtc aaaagggaag ggaactccag tcagaactgg
     5101 caacggtttt accaactgac aaagcttctg gactccatgc atgaggtggt tgagaatctc
     5161 cttacctact gcttccagac atttttggat aagaccatga gtattgaatt cccagagatg
     5221 ttagctgaaa tcatcactaa tcagatacca aaatattcaa atggaaatat caaaaagctt
     5281 ctgtttcatc aaaaatgagc ttctctagct agagtcgatc gacaagctcg agtttctcca
     5341 taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat
     5401 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
     5461 aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcgct atactgaaga
     5521 gcaattctga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     5581 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc
     5641 aaccacaggg ttcccctcgg gatcaaagta ctttgatcca acccctccgc tgctatagtg
     5701 cagtcggctt ctgacgttca gtgcagccgt catctgaaaa cgacatgtcg cacaagtcct
     5761 aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta
     5821 gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg
     5881 ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc
     5941 aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca
     6001 ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct ggcgacgttg
     6061 tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg gacattgccg
     6121 agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg gccgacacca
     6181 ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt
     6241 ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt
     6301 ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag ctgatcgacc
     6361 aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt
     6421 accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct
     6481 tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat gaacgccaag
     6541 aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca ttaccgaaga
     6601 gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc acctctcaac
     6661 cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg cctggccggc
     6721 cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta
     6781 aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata aacaaatacg
     6841 caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc aggcaagacg
     6901 accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt tctgttagtc
     6961 gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta
     7021 accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat cggccggcgc
     7081 gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc cgcgatcaag
     7141 gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg ggccaccgcc
     7201 gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct acaagcggcc
     7261 tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg
     7321 gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc
     7381 actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag
     7441 gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga ggtaaagaga
     7501 aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg
     7561 ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac tttcagttgc
     7621 cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag accattaccg
     7681 agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga ataaatgagt
     7741 agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac
     7801 gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag cggctgggtt
     7861 gcctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgagcggtc
     7921 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
     7981 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
     8041 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
     8101 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
     8161 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
     8221 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
     8281 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
     8341 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
     8401 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
     8461 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
     8521 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
     8581 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
     8641 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
     8701 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
     8761 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
     8821 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
     8881 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
     8941 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
     9001 tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg actctttcct gtg
//
