LOCUS       sequence-386857-        8210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AtTGA1 expression cassette: CaMV35S::TMV::AtTGA1::35sT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8210)
  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 8210)
  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: pEPSW1KN0351
FEATURES             Location/Qualifiers
     source          1..8210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             691..1764
                     /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      1830..2024
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(2260..2282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2368..2508
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2423..2442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2523..2540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2694..3282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2774..2793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3370..4185)
                     /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(3370..4185)
                     /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     4093..4112
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    4618..4642
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4746..5091
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5034..5054
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    6323..6497
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    6518..6542
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             join(7843..8210,1..262)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gcagcgcatt gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat
       61 caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat
      121 tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac
      181 cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga
      241 aattaaatca aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac
      301 acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg
      361 gcagacacgc cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag
      421 ctgaagatgt acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc
      481 gcgcagctac cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa
      541 aggaggcggc atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg
      601 tggaggaacg ggcggttggc caggcgtaag cggctgggtt gcctgccggc cctgcaatgg
      661 cactggaacc cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta
      721 caaatcggcg cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc
      781 cagcggcaac gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat
      841 cgaatccgca aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg
      901 cccaagggcg acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc
      961 gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct
     1021 ggcgaggtga tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc
     1081 ggcatggcca gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa
     1141 tccatgaacc gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac
     1201 gttgcggacg tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg
     1261 gtagaaacct gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc
     1321 aagaacggcc gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc
     1381 gtaaagagcg aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac
     1441 cgcgagatca cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg
     1501 atcgatcccg gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca
     1561 gaagccagat ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag
     1621 aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg
     1681 aaggaggagg cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag
     1741 ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca
     1801 ggggaaaaag gtcgaaaagg actctttcct gtggatagca cgtacattgg gaacccaaag
     1861 ccgtacattg ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg
     1921 tcacacatgt aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct
     1981 ttaaaactta ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc
     2041 acagccgaag tgctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc
     2101 gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc
     2161 aatctaccag ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa
     2221 ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2281 ggtgacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2341 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2401 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2461 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcatccgct
     2521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2641 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2881 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3121 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat
     3361 tctaggtgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     3421 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     3481 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     3541 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     3601 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     3661 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     3721 tcgtgattgc gcctgagcga gtcgaaatac gcgatcgctg ttaaaaggac aattacaaac
     3781 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     3841 atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgcag tggtgagtaa
     3901 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     3961 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     4021 tttcagaaac aactctggcg catcgggctt cccatacaat cggtagattg tcgcacctga
     4081 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     4141 taatcgcggc cttgagcaag acgtttcccg ttgaatatgg ctcataacag aacttattat
     4201 ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga
     4261 actccaattc actgttcctt gcattctaaa accttaaata ccagaaaaca gctttttcaa
     4321 agttgttttc aaagttggcg tataacatag tatcgacgga gccgattttg aaaccgcggt
     4381 gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca
     4441 tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag
     4501 caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc
     4561 tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg
     4621 caggatatat tgtggtgtaa acataacgaa ttgctcttca gcaaggaggt caacatggtg
     4681 gagcacgaca ctctggtcta ctccaaaaat gtcaaagata cagtctcaga agatcaaagg
     4741 gctattgaga cttttcaaca aaggataatt tcgggaaacc tcctcggatt ccattgccca
     4801 gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta caaatgccat
     4861 cattgcgata aaggaaaggc tatcattcaa gatctctctg ccgacagtgg tcccaaagat
     4921 ggacccccac ccacgaggag catcgtggaa aaagaagagg ttccaaccac gtctacaaag
     4981 caagtggatt gatgtgacat ctccactgac gtaagggatg acgcacaatc ccactatcct
     5041 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac acgctcgagt
     5101 ataagagctc atttttacaa caattaccaa caacaacaaa caacaaacaa cattacaatt
     5161 acatttacaa ttatcgatac aatgaattcg acatcgacac attttgtgcc accgagaaga
     5221 gttggtatat acgaacctgt ccatcaattc ggtatgtggg gggagagttt caaaagcaat
     5281 attagcaatg ggactatgaa cacaccaaac cacataataa taccgaataa tcagaaacta
     5341 gacaacaacg tgtcagagga tacttcccat ggaacagcag gaactcctca catgttcgat
     5401 caagaagctt caacgtctag acatcccgat aagatacaaa gacggcttgc tcaaaaccgc
     5461 gaggctgcta ggaaaagtcg cttgcgcaag aaggcttatg ttcagcaact ggaaacaagc
     5521 aggttgaagc taattcaatt agagcaagaa ctcgatcgtg ctagacaaca gggattctat
     5581 gtaggaaacg gaatagatac taattctctc ggtttttcgg aaaccatgaa tccagggatt
     5641 gctgcatttg aaatggaata tggacattgg gttgaagaac agaacagaca gatatgtgaa
     5701 ctaagaacag ttttacacgg acacattaac gatatcgagc ttcgttcgct agtcgaaaac
     5761 gccatgaaac attactttga gcttttccgg atgaaatcgt ctgctgccaa agccgatgtc
     5821 ttcttcgtca tgtcagggat gtggagaact tcagcagaac gattcttctt atggattggc
     5881 ggatttcgac cctccgatct tctcaaggtt cttttgccac attttgatgt cttgacggat
     5941 caacaacttc tagatgtatg caatctaaaa caatcgtgtc agcaagcaga agacgcgttg
     6001 actcaaggta tggagaagct gcaacacacc cttgcggact gcgttgcagc gggacaactc
     6061 ggtgaaggaa gttacattcc tcaggtgaat tctgctatgg atagattaga agctttggtc
     6121 agtttcgtaa atcaggctga tcacttgaga catgaaacat tgcaacaaat gtatcggata
     6181 ttgacaacgc gacaagcggc tcgaggatta ttagctcttg gtgagtattt tcaacggctt
     6241 agagccttga gctcaagttg ggcaactcga catcgtgaac caacgtaggc ttctctagct
     6301 agagtcgatc gacaagctcg agtttctcca taataatgtg tgagtagttc ccagataagg
     6361 gaattagggt tcctataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg
     6421 tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca
     6481 gtactaaaat ccagatcgct atactgaaga gcaattctga caggatatat tggcgggtaa
     6541 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     6601 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
     6661 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
     6721 catctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
     6781 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
     6841 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
     6901 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
     6961 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga
     7021 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
     7081 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
     7141 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
     7201 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
     7261 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
     7321 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
     7381 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
     7441 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
     7501 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
     7561 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
     7621 cgtgttcgag ccgcccgcgc acctctcaac cgtgcggctg catgaaatcc tggccggttt
     7681 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
     7741 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
     7801 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
     7861 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
     7921 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
     7981 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
     8041 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
     8101 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
     8161 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa
//
