LOCUS       sequence-243094-        5583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimeric ETS domain: H2 of Ets-1 in PU.1 scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5583)
  AUTHORS   Albrecht AV, Kim HM, Poon GMK
  TITLE     Mapping interfacial hydration in ETS-family transcription factor 
            complexes with DNA: a chimeric approach.
  JOURNAL   Nucleic Acids Res. 2018 Nov 16;46(20):10577-10588. doi: 
            10.1093/nar/gky894.
  PUBMED    30295801
REFERENCE   2  (bases 1 to 5583)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2018-11-16- 16"; volume: "46"; issue: 
            "20"; pages: "10577-10588"
COMMENT     Sequence Label: pET28b-PU.1-ETS-H2
FEATURES             Location/Qualifiers
     source          1..5583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(179..196)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             524..546
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    561..585
                     /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)."
     primer_bind     complement(585..604)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(586..604)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(688..707)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        913..990
                     /gene="lacI"
                     /label=lacI promoter
     CDS             991..2073
                     /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     complement(1010..1029)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2882..3073
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3067..3089)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3175..3314
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3230..3249
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3329..3346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3500..4088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3580..3599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4199..4218)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4210..5025
                     /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             4210..5025
                     /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     complement(4283..4302)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5117..5572)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5254..5275)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5466..5485
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgc
      181 tcccacgtgg aaccaagtgt ggaggcaggc ggcgctctgc caaaccccca cgccctaata
      241 cttctccact aaactgatag gtcaactttt tctttacctt tttgacctcc ccggttttac
      301 cataattgcg aagggcacgc gccatcttct gataagtcat ttttttgcgg ttaccctttt
      361 gaatccccca acggcgtgcc acctcatcag gatcttttga cgagaactga aaggtcccct
      421 tatctttgtc cacccaccaa atagaatctt tcatgtcgcc actacgcagc aggtcaagta
      481 aaaactgata taagcgaatt tttttcttcg atcccatggt atatctcctt cttaaagtta
      541 aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt
      601 attaatttcg cgggatcgag atctcgatcc tctacgccgg acgcatcgtg gccggcatca
      661 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
      721 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
      781 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
      841 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
      901 gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
      961 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     1021 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     1081 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     1141 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     1201 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     1261 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     1321 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     1381 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     1441 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     1501 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     1561 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     1621 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     1681 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     1741 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     1801 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     1861 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     1921 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     1981 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2041 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     2101 cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2161 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2221 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     2281 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     2341 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     2401 cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     2461 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     2521 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     2581 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     2641 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     2701 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     2761 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     2821 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     2881 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     2941 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     3001 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
     3061 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3121 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     3181 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     3241 cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3301 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3361 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3421 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3601 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3661 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3721 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3841 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3901 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3961 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4021 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4081 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4141 cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa cagtaataca
     4201 aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc
     4261 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     4321 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     4381 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     4441 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     4501 accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     4561 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4621 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     4681 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4741 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     4801 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     4861 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     4921 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     4981 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgag
     5041 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5101 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5161 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5221 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5281 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5341 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5401 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5461 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5521 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     5581 cca
//
