LOCUS       sequence-386946-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector for protein expression of 
            9xHis_AtDREB26_HiBit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  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 6065)
  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: pEPYCdKN0011
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     446..465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     699..716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    731..870
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(796..815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     956..978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(972..1163)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1972..3054)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3016..3035
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3055..3132)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3338..3357
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3441..3460
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3441..3459
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3460..3484
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             3499..3521
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3534..3560
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     protein_bind    3576..3600
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4336..4360)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4370..4387
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4440..4458)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4454..4501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4538..4993
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4625..4644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4835..4856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5085..5900)
                     /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(5085..5900)
                     /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     5808..5827
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5892..5911
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(6022..6065,1..545)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
       61 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      601 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      661 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      721 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      781 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
      841 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
      901 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
      961 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     1021 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     1081 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     1141 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     1201 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     1261 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     1321 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     1381 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     1441 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     1501 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     1561 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     1621 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     1681 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     1741 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1801 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1861 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     1921 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     1981 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2041 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     2101 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     2161 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     2221 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     2281 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     2341 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     2401 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     2461 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     2521 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     2581 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     2641 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     2701 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     2761 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2821 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     2881 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     2941 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     3001 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     3061 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     3121 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     3181 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     3241 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     3301 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     3361 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     3421 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     3481 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg aaacaccacc
     3541 accaccacca ccaccaccac ggtggtctcg aatcaacaag tttgtacaaa aaagcaggct
     3601 tcatggtgaa acaagaacgc aagatccaaa ccagcagcac aaaaaaggaa atgcctttgt
     3661 catcatcacc atcttcttct tcttcttcat cttcttcctc gtcttcgtct tcgtgtaaga
     3721 acaagaacaa gaagagtaag attaagaagt acaaaggagt gaggatgaga agttggggat
     3781 catgggtttc tgagattagg gcaccaaatc aaaagacaag gatttggtta ggttcttact
     3841 caacagctga agcagctgct agagcttacg atgttgcact cttatgtctc aaaggccctc
     3901 aagccaatct caacttccct acttcttctt cttctcatca tcttcttgat aatctcttag
     3961 atgaaaatac ccttttgtcc cccaaatcca tccaaagagt agctgctcaa gctgccaact
     4021 catttaacca ttttgcccct acttcatcag ccgtctcgtc accgtccgat catgatcatc
     4081 accatgatga tgggatgcaa tctttgatgg gatcttttgt ggacaatcat gtgtctttga
     4141 tggattcaac atcttcatgg tatgatgatc ataatgggat gttcttgttt gataatggag
     4201 ctccattcaa ttactctcct caactaaact cgacgacgat gctcgatgaa tacttctacg
     4261 aagatgctga cattccgctt tggagtttca atggttcggt gagcggctgg cggctgttca
     4321 agaagattag ctgaaaccca gctttcttgt acaaagtggt tgattcgagc accaccacca
     4381 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     4441 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     4501 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     4561 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4621 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4681 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4741 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4801 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4861 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4921 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4981 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5041 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     5101 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     5161 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     5221 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     5281 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     5341 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     5401 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     5461 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     5521 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     5581 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     5641 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     5701 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     5761 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     5821 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     5881 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     5941 attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6001 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6061 aacaa
//
