LOCUS       sequence-258615-        6742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ETS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6742)
  AUTHORS   Selvaraj N, Kedage V, Hollenhorst PC
  TITLE     Comparison of MAPK specificity across the ETS transcription factor 
            family identifies a high-affinity ERK interaction required for ERG 
            function in prostate cells.
  JOURNAL   Cell Commun Signal. 2015 Feb 19;13:12. doi: 
            10.1186/s12964-015-0089-7.
  PUBMED    25885538
REFERENCE   2  (bases 1 to 6742)
  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"; journalName: "Cell Commun
            Signal."; date: "2015-02-19"; volume: "13:12. doi"; pages: " 
            10.1186/s12964-015-0089-7"
COMMENT     Sequence Label: ETS2_pet28a
FEATURES             Location/Qualifiers
     source          1..6742
                     /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(1620..1637)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1647..1664)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1683..1705
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1720..1744
                     /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(1744..1763)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1745..1763)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1847..1866)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2072..2149
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2150..3232
                     /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(2169..2188)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4041..4232
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4226..4248)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4334..4473
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4389..4408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4488..4505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4659..5247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4739..4758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5358..5377)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5369..6184
                     /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             5369..6184
                     /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(5442..5461)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6276..6731)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6413..6434)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6625..6644
                     /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 gcaagcttgt
      181 cgacggagct cgaattcgga tcctcatcag tcctccgtgt cgggctggac gcccaggatg
      241 gcgtgcagtt cctcgggcgt gaaccccagc aagttctgga ggtcgcacac gaagcggtac
      301 acgtagcgct tccccgacgt cttgtggatg atgttcttgt cataatagta gcgtaagccc
      361 cggctcagct tctcgtagtt catcttgggc ttatttttcc tctttcccca ccggcgggcc
      421 acctcatcgg ggtcggcgag cttaaactcc catccgtctc cagtccagct gatgaatgac
      481 tggcaggatt tgtctgatag cagctccagg agaaactgcc acagctgaat aggtccactt
      541 cctgtgaagc cggccagcac agctgcaggt ataactggtt tgccttgctc caccgggtca
      601 ctcctctctt ggatgtaatc cttgaaagac atggttggct tattgaggca gagagactgg
      661 ctgcagtcat cttcgaagct ctcgaaggaa ggaacccgtt gcacatccag caaggacgac
      721 tggctgttcc aggactggag gagggagtct gagctctcga agctgtccgc accgttctca
      781 ggggagtcgt ggtctttggg cgtcccagaa ttgttggtga gcaaattcaa gttgctgcct
      841 gggaagtcct gactgacaga gcagtaggtg acgctgacgg agctgagccg agacttgggg
      901 aacatctgaa actcctgctc agagctgagt acgctgggtg tggaggccgg acacatgctg
      961 tccaggaggc cgcctttggg gtaattctgt gtctgcattc catagggcgc ctgctctgtg
     1021 ccaaaaccta atgtattgct gttaatccaa tgaggaacgg aggtgaggtg tgaattttct
     1081 tcatattgat cttctgtctt ttcttggttt tctttgatca tttgctccag atgttcccag
     1141 agaatgtcac ccacaaagtc aggtgccagc tccagaaagc gttccttgcc aaggttacac
     1201 agcatctggc cattcatgcc gaacctctgc agattcacgt tcaccagact gaactcattg
     1261 gtggcccaga gaagccactg gcatacctgt tgctcactcc acagccaggg gttctttgga
     1321 atgcccaggc gccgctgttc ctttttgaag ccactgaagg tagcttttaa ggcttgactc
     1381 atcacagcct tgctgcacgg ggttaacaaa ggcaattcac agttggcgga gtcatgagaa
     1441 atggaatcca agcctgttgg cacttcttgc agtgtttgct cttcatttag agaaggaaaa
     1501 acagcaaaca gggacccatc aaaggtgtca aaggctggct ggcgcttgag tgtccctctg
     1561 taactgttag ccacaggggc tacctggtcc atattcttga ttccgaaatc attcatatgg
     1621 ctgccgcgcg gcaccaggcc gctgctgtga tgatgatgat gatggctgct gcccatggta
     1681 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
     1741 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga
     1801 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     1861 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     1921 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     1981 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     2041 gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg
     2101 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     2161 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     2221 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     2281 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     2341 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     2401 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     2461 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     2521 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     2581 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     2641 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     2701 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     2761 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     2821 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     2881 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     2941 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     3001 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     3061 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     3121 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3181 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3241 caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag
     3301 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     3361 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     3421 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     3481 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     3541 agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt
     3601 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     3661 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     3721 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     3781 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     3841 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     3901 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     3961 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     4021 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     4081 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     4141 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     4201 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4261 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4321 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     4381 taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg
     4441 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     4501 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4561 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4621 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4681 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4741 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4801 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4861 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4921 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4981 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5041 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5101 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5161 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5221 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5281 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc
     5341 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
     5401 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     5461 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     5521 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     5581 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     5641 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     5701 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     5761 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     5821 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     5881 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5941 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     6001 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     6061 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     6121 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     6181 ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6241 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     6301 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6361 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6421 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6481 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6541 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6601 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6661 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6721 acagggcgcg tcccattcgc ca
//
