LOCUS       sequence-387006-        9020 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 9020)
  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 9020)
  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: pEPSW1KN0113
FEATURES             Location/Qualifiers
     source          1..9020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(99..121)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    207..347
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     262..281
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(362..379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(533..1121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(613..632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1209..2024)
                     /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(1209..2024)
                     /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     1932..1951
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2457..2481
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2585..2930
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2873..2893
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     regulatory      3401..3410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4972..5146
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5167..5191
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6492..7121
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7550..8623
                     /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      8689..8883
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 atctaccagg gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag
       61 gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      121 gtgacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      181 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
      241 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
      301 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcatccgctt
      361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      961 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt
     1201 ctaggtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     1261 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     1321 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     1381 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     1441 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     1501 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     1561 cgtgattgcg cctgagcgag tcgaaatacg cgatcgctgt taaaaggaca attacaaaca
     1621 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     1681 tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt ggtgagtaac
     1741 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1801 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     1861 ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt cgcacctgat
     1921 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     1981 aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacaga acttattatt
     2041 tccttcctct tttctacagt atttaaagat accccaagaa gctaattata acaagacgaa
     2101 ctccaattca ctgttccttg cattctaaaa ccttaaatac cagaaaacag ctttttcaaa
     2161 gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga aaccgcggtg
     2221 atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc gcgagatcat
     2281 ccgtgtttca aacccggcag cttagttgcc gttcttccga atagcatcgg taacatgagc
     2341 aaagtctgcc gccttacaac ggctctcccg ctgacgccgt cccggactga tgggctgcct
     2401 gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc
     2461 aggatatatt gtggtgtaaa cataacgaat tgctcttcag caaggaggtc aacatggtgg
     2521 agcacgacac tctggtctac tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg
     2581 ctattgagac ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag
     2641 ctatctgtca cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc
     2701 attgcgataa aggaaaggct atcattcaag atctctctgc cgacagtggt cccaaagatg
     2761 gacccccacc cacgaggagc atcgtggaaa aagaagaggt tccaaccacg tctacaaagc
     2821 aagtggattg atgtgacatc tccactgacg taagggatga cgcacaatcc cactatcctt
     2881 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca cgctcgagta
     2941 taagagctca tttttacaac aattaccaac aacaacaaac aacaaacaac attacaatta
     3001 catttacaat tatcgataca atggcaaatc gcggaggtga atatctgtac gatgagttat
     3061 ggaaattatg cgcgggacct cttgttgatg ttcctcaagc tcaagaaaga gtttattatt
     3121 ttcctcaagg tcacatggaa caactcgaag cgtcaacgca acaagtcgac ttaaatacga
     3181 tgaagcctct ttttgttctt cctcctaaga ttctctgcaa tgttatgaac gttagtcttc
     3241 aggcggagaa agatacggat gaggtctatg ctcagattac tttgatccct gttggaactg
     3301 aagttgatga acctatgagt cctgatccct ctcctcctga gttgcaaagg ccgaaagttc
     3361 actctttcag caaggttttg acagcgtctg atacaagcac ccatggtggc ttttctgttc
     3421 taaggaaaca tgccacggaa tgtcttcctc cgctggatat gactcagcaa accccgaccc
     3481 aggagttagt agccgaagat gtgcacggtt atcagtggaa attcaagcat atttttagag
     3541 gccaaccacg gaggcatcta ttgacgacag ggtggagcac ctttgttaca tcaaagagat
     3601 tggttgctgg ggacaccttt gtattcctga gaggggagaa cggagagttg cgagttggag
     3661 tcagacgtgc taatcttcaa cagagcagta tgccttcatc cgttatatca agtcatagta
     3721 tgcatctggg agtgcttgct actgcacgcc atgctactca aacgaaaact atgttcattg
     3781 tatactataa accaaggaca agccaattca tcattagctt gaacaaatat ctagaagcca
     3841 tgagcaataa gttctctgta gggatgagat ttaagatgcg ttttgaggga gaggattccc
     3901 ctgaaagaag atattctggc acggttattg gtgtgaaaga ctgctcccct cactggaaag
     3961 actcaaaatg gcgatgctta gaagttcatt gggatgagcc tgcatcgatt tcaagaccca
     4021 ataaggtttc accatgggaa attgaaccgt ttgtaaattc agaaaacgtt cccaaatcag
     4081 ttatgctgaa gaacaaaagg ccccgtcaag ttagtgaagt atctgcactt gatgtaggca
     4141 taacagcttc aaacctttgg agctctgttt tgacgcaacc ccatgagttt gcacaatcgt
     4201 gcatcacctc acagtggagt tctcctcagc aatgtcatcg tgatgcaaat gaggatgcta
     4261 agaaatctga ctggctaaat aactcttact ctgtgtcaaa tgtagcaaaa gactcaacac
     4321 tgaacgacca aatggtttcc ccagtcgagc agaagaagcc tgagacaacc gctaattata
     4381 gattatttgg aattgatctg atgagttcct ccctagcggt tcctgaggag aaaactgcac
     4441 ccatgcgacc aatcaacata tccaaaccga ctatggacag ccactcagac ccaaaatcag
     4501 agatttcaaa agtatcagaa gagaaaaagc aggaacctgc ggagggatca ccaaaagagg
     4561 tccaaagcaa gcaaagcagt tctacaagaa gccgtaccaa ggtgcagatg caaggcgtac
     4621 ctgtgggcag ggctgtggat ttaaatgcgc taaaggggta caacgagctc atagatgaca
     4681 ttgagaagct gtttgacata aaaggggaac tgcggagtcg caatcaatgg gaaatagtgt
     4741 tcacagacga tgagggagat atgatgcttg tcggtgatga cccatggcct gagttctgca
     4801 acatggtgaa gagaatattc atatggtcga aagaggaagt gaagaaaatg acgcctggga
     4861 accaactccg gatgctgtta agggaagttg aaacaacact aacaacaact tccaaaacag
     4921 ataatcattc caactaagct tctctagcta gagtcgatcg acaagctcga gtttctccat
     4981 aataatgtgt gagtagttcc cagataaggg aattagggtt cctatagggt ttcgctcatg
     5041 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
     5101 aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatcgcta tactgaagag
     5161 caattctgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa
     5221 tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca
     5281 accacagggt tcccctcggg atcaaagtac tttgatccaa cccctccgct gctatagtgc
     5341 agtcggcttc tgacgttcag tgcagccgtc atctgaaaac gacatgtcgc acaagtccta
     5401 agttacgcga caggctgccg ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag
     5461 tcgcataaag tagaatactt gcgactagaa ccggagacat tacgccatga acaagagcgc
     5521 cgccgctggc ctgctgggct atgcccgcgt cagcaccgac gaccaggact tgaccaacca
     5581 acgggccgaa ctgcacgcgg ccggctgcac caagctgttt tccgagaaga tcaccggcac
     5641 caggcgcgac cgcccggagc tggccaggat gcttgaccac ctacgccctg gcgacgttgt
     5701 gacagtgacc aggctagacc gcctggcccg cagcacccgc gacctactgg acattgccga
     5761 gcgcatccag gaggccggcg cgggcctgcg tagcctggca gagccgtggg ccgacaccac
     5821 cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc attgccgagt tcgagcgttc
     5881 cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt
     5941 tggcccccgc cctaccctca ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca
     6001 ggaaggccgc accgtgaaag aggcggctgc actgcttggc gtgcatcgct cgaccctgta
     6061 ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag gccaggcggc gcggtgcctt
     6121 ccgtgaggac gcattgaccg aggccgacgc cctggcggcc gccgagaatg aacgccaaga
     6181 ggaacaagca tgaaaccgca ccaggacggc caggacgaac cgtttttcat taccgaagag
     6241 atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca cctctcaacc
     6301 gtgcggctgc atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc
     6361 agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa
     6421 aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa acaaatacgc
     6481 aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca ggcaagacga
     6541 ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg
     6601 attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat caaccgctaa
     6661 ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc ggccggcgcg
     6721 acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg
     6781 cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacatatgg gccaccgccg
     6841 acctggtgga gctggttaag cagcgcattg aggtcacgga tggaaggcta caagcggcct
     6901 ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg
     6961 ccgggtacga gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc tacccaggca
     7021 ctgccgccgc cggcacaacc gttcttgaat cagaacccga gggcgacgct gcccgcgagg
     7081 tccaggcgct ggccgctgaa attaaatcaa aactcatttg agttaatgag gtaaagagaa
     7141 aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc
     7201 tgcaacgttg gccagcctgg cagacacgcc agccatgaag cgggtcaact ttcagttgcc
     7261 ggcggaggat cacaccaagc tgaagatgta cgcggtacgc caaggcaaga ccattaccga
     7321 gctgctatct gaatacatcg cgcagctacc agagtaaatg agcaaatgaa taaatgagta
     7381 gatgaatttt agcggctaaa ggaggcggca tggaaaatca agaacaacca ggcaccgacg
     7441 ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg
     7501 cctgccggcc ctgcaatggc actggaaccc ccaagcccga ggaatcggcg tgagcggtcg
     7561 caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt
     7621 gaaggccgcg caggccgccc agcggcaacg catcgaggca gaagcacgcc ccggtgaatc
     7681 gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg caaccgccgg cagccggtgc
     7741 gccgtcgatt aggaagccgc ccaagggcga cgagcaacca gattttttcg ttccgatgct
     7801 ctatgacgtg ggcacccgcg atagtcgcag catcatggac gtggccgttt tccgtctgtc
     7861 gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag cttccagacg ggcacgtaga
     7921 ggtttccgca gggccggccg gcatggccag tgtgtgggat tacgacctgg tactgatggc
     7981 ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa gggaagggag acaagcccgg
     8041 ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc tgccggcgag ccgatggcgg
     8101 aaagcagaaa gacgacctgg tagaaacctg cattcggtta aacaccacgc acgttgccat
     8161 gcagcgtacg aagaaggcca agaacggccg cctggtgacg gtatccgagg gtgaagcctt
     8221 gattagccgc tacaagatcg taaagagcga aaccgggcgg ccggagtaca tcgagatcga
     8281 gctagctgat tggatgtacc gcgagatcac agaaggcaag aacccggacg tgctgacggt
     8341 tcaccccgat tactttttga tcgatcccgg catcggccgt tttctctacc gcctggcacg
     8401 ccgcgccgca ggcaaggcag aagccagatg gttgttcaag acgatctacg aacgcagtgg
     8461 cagcgccgga gagttcaaga agttctgttt caccgtgcgc aagctgatcg ggtcaaatga
     8521 cctgccggag tacgatttga aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg
     8581 ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg agcagatgct
     8641 agggcaaatt gccctagcag gggaaaaagg tcgaaaagga ctctttcctg tggatagcac
     8701 gtacattggg aacccaaagc cgtacattgg gaaccggaac ccgtacattg ggaacccaaa
     8761 gccgtacatt gggaaccggt cacacatgta agtgactgat ataaaagaga aaaaaggcga
     8821 tttttccgcc taaaactctt taaaacttat taaaactctt aaaacccgcc tggcctgtgc
     8881 ataactgtct ggccagcgca cagccgaagt gctgcaaaaa gcgcctaccc ttcggtcgct
     8941 gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat
     9001 ggctggccta cggccaggca
//
