LOCUS       sequence-387319-        7796 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 7796)
  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 7796)
  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: pEPSW1KN0030
FEATURES             Location/Qualifiers
     source          1..7796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      176..370
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(606..628)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    714..854
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     769..788
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(869..886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1040..1628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1120..1139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1716..2531)
                     /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(1716..2531)
                     /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     2439..2458
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2964..2988
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3092..3437
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3380..3400
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    4255..4429
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4450..4474
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5775..6404
                     /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(6833..7796,1..110)
                     /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 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag
       61 tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc
      121 agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggactc tttcctgtgg
      181 atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga
      241 acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa
      301 aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg
      361 cctgtgcata actgtctggc cagcgcacag ccgaagtgct gcaaaaagcg cctacccttc
      421 ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct
      481 caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc
      541 cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt
      601 gaaaacctct gacacatgca gctcccggtg acggtcacag cttgtctgta agcggatgcc
      661 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
      721 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
      781 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
      841 aataccgcat caggcgctca tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      901 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      961 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1021 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1081 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1141 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1201 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1261 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1321 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1381 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1441 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     1501 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1561 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1621 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1681 cacgttaagg gattttggtc atgcattcta ggtgattaga aaaactcatc gagcatcaaa
     1741 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1801 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1861 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1921 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1981 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2041 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagtcg aaatacgcga
     2101 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2161 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2221 ttccctggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2281 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2341 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2401 tacaatcggt agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2461 tataaatcag catccatgtt ggaatttaat cgcggccttg agcaagacgt ttcccgttga
     2521 atatggctca taacagaact tattatttcc ttcctctttt ctacagtatt taaagatacc
     2581 ccaagaagct aattataaca agacgaactc caattcactg ttccttgcat tctaaaacct
     2641 taaataccag aaaacagctt tttcaaagtt gttttcaaag ttggcgtata acatagtatc
     2701 gacggagccg attttgaaac cgcggtgatc acaggcagca acgctctgtc atcgttacaa
     2761 tcaacatgct accctccgcg agatcatccg tgtttcaaac ccggcagctt agttgccgtt
     2821 cttccgaata gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc tctcccgctg
     2881 acgccgtccc ggactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg
     2941 tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacat aacgaattgc
     3001 tcttcagcaa ggaggtcaac atggtggagc acgacactct ggtctactcc aaaaatgtca
     3061 aagatacagt ctcagaagat caaagggcta ttgagacttt tcaacaaagg ataatttcgg
     3121 gaaacctcct cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa
     3181 aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatc
     3241 tctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
     3301 aagaggttcc aaccacgtct acaaagcaag tggattgatg tgacatctcc actgacgtaa
     3361 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
     3421 ttcatttgga gaggacacgc tcgagtataa gagctcattt ttacaacaat taccaacaac
     3481 aacaaacaac aaacaacatt acaattacat ttacaattat cgatacaatg gtgaaacaag
     3541 aacgcaagat ccaaaccagc agcacaaaaa aggaaatgcc tttgtcatca tcaccatctt
     3601 cttcttcttc ttcatcttct tcctcgtctt cgtcttcgtg taagaacaag aacaagaaga
     3661 gtaagattaa gaagtacaaa ggagtgagga tgagaagttg gggatcatgg gtttctgaga
     3721 ttagggcacc aaatcaaaag acaaggattt ggttaggttc ttactcaaca gctgaagcag
     3781 ctgctagagc ttacgatgtt gcactcttat gtctcaaagg ccctcaagcc aatctcaact
     3841 tccctacttc ttcttcttct catcatcttc ttgataatct cttagatgaa aatacccttt
     3901 tgtcccccaa atccatccaa agagtagctg ctcaagctgc caactcattt aaccattttg
     3961 cccctacttc atcagccgtc tcgtcaccgt ccgatcatga tcatcaccat gatgatggga
     4021 tgcaatcttt gatgggatct tttgtggaca atcatgtgtc tttgatggat tcaacatctt
     4081 catggtatga tgatcataat gggatgttct tgtttgataa tggagctcca ttcaattact
     4141 ctcctcaact aaactcgacg acgatgctcg atgaatactt ctacgaagat gctgacattc
     4201 cgctttggag tttcaattaa gcttctctag ctagagtcga tcgacaagct cgagtttctc
     4261 cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc
     4321 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     4381 ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcg ctatactgaa
     4441 gagcaattct gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     4501 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
     4561 ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc gctgctatag
     4621 tgcagtcggc ttctgacgtt cagtgcagcc gtcatctgaa aacgacatgt cgcacaagtc
     4681 ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt
     4741 tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca tgaacaagag
     4801 cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg acttgaccaa
     4861 ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga agatcaccgg
     4921 caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc ctggcgacgt
     4981 tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac tggacattgc
     5041 cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt gggccgacac
     5101 caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg
     5161 ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa
     5221 gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg agctgatcga
     5281 ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct
     5341 gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc
     5401 cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga atgaacgcca
     5461 agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt cattaccgaa
     5521 gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc gcacctctca
     5581 accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg
     5641 gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag
     5701 taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata
     5761 cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga
     5821 cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag
     5881 tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc
     5941 taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc
     6001 gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca
     6061 aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg
     6121 ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg
     6181 cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc
     6241 tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag
     6301 gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg
     6361 aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga
     6421 gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa
     6481 ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt
     6541 gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac
     6601 cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga
     6661 gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg
     6721 acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg
     6781 ttgcctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg
     6841 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
     6901 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
     6961 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
     7021 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
     7081 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
     7141 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
     7201 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
     7261 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
     7321 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
     7381 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
     7441 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
     7501 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
     7561 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
     7621 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
     7681 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
     7741 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggt
//
