LOCUS       sequence-322043-        8548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expression of the transcriptional activator 
            protein CUP2 in C-terminal fusion with GAL4 activation domain 
            (GAL4AD). Binds to specific DNA operator in the presence of copper 
            ions and promotes transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8548)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 8548)
  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"
COMMENT     Sequence Label: pEGB2_alpha2 P35S:CUP2:GAL4AD:T35S (GB1039)
FEATURES             Location/Qualifiers
     source          1..8548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             3523..4152
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4581..5654
                     /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      5720..5914
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6150..6172)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6258..6398
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6313..6332
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6413..6430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6584..7172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6664..6683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7260..8075)
                     /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(7260..8075)
                     /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     7983..8002
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8508..8530
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcccctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
     1021 tattctagtc gaaatgatgg tcgtaattaa cggggtcaaa tatgcctgtg aaacgtgtat
     1081 caggggtcac agggcggcgc agtgtactca cactgatggt ccgctacaga tgatcagacg
     1141 caagggaaga ccatcgacca catgtggcca ttgtaaagag ctgagaagaa ccaagaactt
     1201 caacccatcc ggtgggtgca tgtgtgcctc tgcacgacgg ccagctgttg gcagcaagga
     1261 agatgaaaca cgatgtcgtt gtgatgaggg tgaaccttgt aaatgtcata ccaagaggaa
     1321 aagcagccgg aaatcaaagg gagggtcatg ccacagaagg gcaaatgatg aagcagcgca
     1381 tgtcaatggt cttggtattg cagatctgga cgttcttttg ggcctaaatg gtcgctcgtc
     1441 ggatgtagac atgacaacca cattgccgag tttgaagcca cctctgcaaa acggagaaat
     1501 taaggccgac agcattgaca atcttgattt ggcttccctc gatccgcttg agcaaagccc
     1561 tagtatatct atggaacctg ttagtatcaa tgaaacagga agcgcatata caactacgaa
     1621 cacagcacta aacgatattg acattccatt ctccatcaat gagttgaacg agctatacaa
     1681 acaagtatct tcgcataact cacattcaca gccaacaaat tctcaagcgc tttcacaacc
     1741 aattgcctcc tctaacgttc atgataactt catgaataat gaaatcacgg ctagtaaaat
     1801 tgatgatggt aataattcaa aaccactgtc acctggttgg acggaccaaa ctgcgtataa
     1861 cgcgtttgga atcactacag ggatgtttaa taccactaca atggatgatg tatataacta
     1921 tctattcgat gatgaagata ccccaccaaa cccaaaaaaa gagtgagctt cggccatgct
     1981 agagtccgca aaaatcacca gtctctctct acaaatctat ctctctctat ttttctccag
     2041 aataatgtgt gagtagttcc cagataaggg aattagggtt cttatagggt ttcgctcatg
     2101 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
     2161 aaatttctaa ttcctaaaac caaaatccag tgacctcgct tgagacgaag cttgacgtaa
     2221 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     2281 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
     2341 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
     2401 catctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
     2461 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
     2521 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
     2581 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
     2641 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcctggag ctggccagga
     2701 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
     2761 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
     2821 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
     2881 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
     2941 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
     3001 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
     3061 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
     3121 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
     3181 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
     3241 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
     3301 cgtgttcgag ccgcccgcgc acctctcaac cgtgcggctg catgaaatcc tggccggttt
     3361 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
     3421 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
     3481 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
     3541 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
     3601 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
     3661 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
     3721 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
     3781 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
     3841 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
     3901 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
     3961 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
     4021 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
     4081 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
     4141 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
     4201 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
     4261 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
     4321 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
     4381 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
     4441 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
     4501 ggcggttggc caggcgtaag cggctgggtt gcctgccggc cctgcaatgg cactggaacc
     4561 cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg
     4621 cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac
     4681 gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca
     4741 aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg
     4801 acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca
     4861 gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga
     4921 tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca
     4981 gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc
     5041 gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg
     5101 tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct
     5161 gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc
     5221 gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg
     5281 aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca
     5341 cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg
     5401 gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat
     5461 ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt
     5521 tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg
     5581 cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat
     5641 ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag
     5701 gtcgaaaagg actctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg
     5761 ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt
     5821 aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta
     5881 ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag
     5941 agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc
     6001 ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag
     6061 ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc
     6121 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggtgacggtc
     6181 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6241 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     6301 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6361 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     6421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7021 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtgat
     7261 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     7321 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     7381 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     7441 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     7501 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     7561 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     7621 gcctgagcga gtcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     7681 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     7741 tcttctaata cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca
     7801 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     7861 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     7921 aactctggcg catcgggctt cccatacaat cggtagattg tcgcacctga ttgcccgaca
     7981 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     8041 cttgagcaag acgtttcccg ttgaatatgg ctcataacag aacttattat ttccttcctc
     8101 ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga actccaattc
     8161 actgttcctt gcattctaaa accttaaata ccagaaaaca gctttttcaa agttgttttc
     8221 aaagttggcg tataacatag tatcgacgga gccgattttg aaaccgcggt gatcacaggc
     8281 agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc
     8341 aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc
     8401 cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt
     8461 ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat
     8521 tgtggtgtaa caagcttcgt ctcagtca
//
