LOCUS       sequence-387004-        7865 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 7865)
  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 7865)
  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: pEPSW1KN0031
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      213..407
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(643..665)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    751..891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     806..825
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(906..923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1077..1665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1157..1176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1753..2568)
                     /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(1753..2568)
                     /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     2476..2495
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3001..3025
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3129..3474
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3417..3437
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    4361..4535
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4556..4580
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5881..6510
                     /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(6939..7865,1..147)
                     /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 aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat
       61 ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc
      121 gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta
      181 gcaggggaaa aaggtcgaaa aggactcttt cctgtggata gcacgtacat tgggaaccca
      241 aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac
      301 cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac
      361 tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag
      421 cgcacagccg aagtgctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc
      481 gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca
      541 ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat
      601 caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
      661 cccggtgacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      721 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
      781 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
      841 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcatcc
      901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1501 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1741 cattctaggt gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     1801 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1861 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1921 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1981 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     2041 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2101 cattcgtgat tgcgcctgag cgagtcgaaa tacgcgatcg ctgttaaaag gacaattaca
     2161 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2221 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag
     2281 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2341 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2401 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc
     2461 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2521 atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcataa cagaacttat
     2581 tatttccttc ctcttttcta cagtatttaa agatacccca agaagctaat tataacaaga
     2641 cgaactccaa ttcactgttc cttgcattct aaaaccttaa ataccagaaa acagcttttt
     2701 caaagttgtt ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc
     2761 ggtgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga
     2821 tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat
     2881 gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct
     2941 gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg
     3001 tggcaggata tattgtggtg taaacataac gaattgctct tcagcaagga ggtcaacatg
     3061 gtggagcacg acactctggt ctactccaaa aatgtcaaag atacagtctc agaagatcaa
     3121 agggctattg agacttttca acaaaggata atttcgggaa acctcctcgg attccattgc
     3181 ccagctatct gtcacttcat cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc
     3241 catcattgcg ataaaggaaa ggctatcatt caagatctct ctgccgacag tggtcccaaa
     3301 gatggacccc cacccacgag gagcatcgtg gaaaaagaag aggttccaac cacgtctaca
     3361 aagcaagtgg attgatgtga catctccact gacgtaaggg atgacgcaca atcccactat
     3421 ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag gacacgctcg
     3481 agtataagag ctcattttta caacaattac caacaacaac aaacaacaaa caacattaca
     3541 attacattta caattatcga tacaatgatg aaatctgggg ctgatttgca atttccacca
     3601 ggatttagat ttcatcctac ggatgaggag ctagtcctca tgtatctctg tcgtaaatgc
     3661 gcgtcgcagc cgatccctgc tccgattatc accgaactcg atttgtaccg atatgatcct
     3721 tgggaccttc ccgacatggc tttgtacggt gaaaaggagt ggtatttttt ctcaccaaga
     3781 gatcgaaagt atccaaacgg ttcaagaccc aaccgtgcag ctggtactgg atattggaaa
     3841 gctaccggag ctgataaacc aataggtcgt cctaaaccgg ttggtattaa gaaggctcta
     3901 gtgttttact cgggaaaacc tccaaatgga gagaaaacca attggattat gcacgaatac
     3961 cggctcgctg acgttgaccg gtcggttcgt aagaaaaaca gtctaagatt ggacgattgg
     4021 gtattgtgtc gtatatataa caagaaaggt gtcatcgaga agcgacgaag cgatatcgag
     4081 gacgggttaa agcctgtgac tgacacgtgt ccaccggaat ctgtggcgag attgatctcc
     4141 ggctcggagc aagcggtgtc accggaattc acgtgtagca acggtcggtt gagtaatgcc
     4201 cttgattttc cgtttaatta cgtagatgcc atcgccgata acgagattgt gtcacggcta
     4261 ttgggcggga atcagatgtg gtcgacgacg cttgatccac ttgtggttag gcagggaact
     4321 ttctgagctt ctctagctag agtcgatcga caagctcgag tttctccata ataatgtgtg
     4381 agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata
     4441 taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat
     4501 tcctaaaacc aaaatccagt actaaaatcc agatcgctat actgaagagc aattctgaca
     4561 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     4621 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     4681 cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct
     4741 gacgttcagt gcagccgtca tctgaaaacg acatgtcgca caagtcctaa gttacgcgac
     4801 aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt
     4861 agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc
     4921 tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac
     4981 tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc
     5041 gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca
     5101 ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg
     5161 aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg
     5221 gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg
     5281 accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc
     5341 ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca
     5401 ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg
     5461 agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg
     5521 cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat
     5581 gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga
     5641 gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ctctcaaccg tgcggctgca
     5701 tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca gcttggccgc
     5761 tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg
     5821 tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc
     5881 atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc
     5941 catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc
     6001 cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc
     6061 atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg
     6121 atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc
     6181 gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag
     6241 ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg
     6301 cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag
     6361 ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc
     6421 ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg
     6481 gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa
     6541 gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg
     6601 ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc
     6661 acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg
     6721 aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta
     6781 gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc
     6841 cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc ctgccggccc
     6901 tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc aaaccatccg
     6961 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc
     7021 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
     7081 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
     7141 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
     7201 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
     7261 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
     7321 ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
     7381 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
     7441 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
     7501 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
     7561 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
     7621 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
     7681 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
     7741 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
     7801 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
     7861 agttc
//
