LOCUS       sequence-261771-        6863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ETV3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6863)
  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 6863)
  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: ETV3_pet28a
FEATURES             Location/Qualifiers
     source          1..6863
                     /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(1741..1758)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1768..1785)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1804..1826
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1841..1865
                     /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(1865..1884)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1866..1884)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1968..1987)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2193..2270
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2271..3353
                     /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(2290..2309)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4162..4353
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4347..4369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4455..4594
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4510..4529
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4609..4626)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4780..5368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4860..4879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5479..5498)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5490..6305
                     /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             5490..6305
                     /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(5563..5582)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6397..6852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6534..6555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6746..6765
                     /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 cgaattcact aagcatcagc agctgctgtt gccaagccct ggggtcctga
      241 cccattccag agaaacttgc cactcttgct cagctctcgg gcttcagggt catcattcca
      301 gcgccgcttc aaccgaagct tggggggcat cagtgcatct tctttcttct caggtgcact
      361 gggctctttg ccaggcctat cctcactgtc ttcagtcacc tccagaggtt ctccacttgg
      421 ggtgctaatg ggcacagagg gccagatggg tggtgcagca ggacgggcaa agatggtgcc
      481 tttttcctct tcaatggtcc tgcttggcac agtgccctct tcttgagtgt gctcctcctt
      541 ctctcgtgcc gactgcctga ggctctcagg atccttttca gaggcaggtt ccaccttgat
      601 tcttggggga atagaagcca tggtgggcgt ggtgactggt gctgactcct cgctgctctc
      661 aaccctctcc cggttcttcc gcccaactgg tgggggctgc agcttgatgg agaactgagt
      721 tgactcctca ggatgcattt ggcactgcag tggtggaacc atgagccctg ggtaacgggg
      781 aaaagtccga ggactcagat ggtagttgaa cacagaacaa gcttgagaat gaaggtagtg
      841 tttcatttcc tcagggttga aggagaagca gctgctgctg gtgaaagggc tcaggcctgg
      901 tgatgggcta taggagaaga tggtgggagt cagggagagg gctggtgaaa tggggacatt
      961 aaggacacct ccgcggccag ggattggaga gacagcgaag ggactgtggg ggtcagggta
     1021 catccctggc ctagcaaaca gaggaagcat tatgtcaggc ttgcgtttct gatggccaat
     1081 ccctcctcca ccaatggcat tcctggagga cacgggatcc acaccccggc gccagtcagc
     1141 agctgagcca tcctccagct ctgaaagctc agtctttcta tcagtaccat tactggactc
     1201 ctggccagaa gcagttaggg agctagcaga gaaccgtccc ggctgcacat cattggttgg
     1261 agaatgggtg tccagaggtg ggaaatggaa ccgggaagag gctgttggca ctggtggtgc
     1321 actctgagga accacaccac ttgaccgaat gttgatgaat gggtagttgg gcatcaccag
     1381 cttgttgaag ttaaatttat aggtaaatct tttccctttt gttttatgaa ggatcctctt
     1441 gttgtaatag tatctgaggg cccggctcag cttgtcataa ttcatctgtg gtttgcattt
     1501 cctgcggccc cagaggcggg ccacctcatc tggatccttg atgacaaatt ccccgtactc
     1561 tccctgctgc caggcgatga catggcggaa ctcttccttc tgcagcagct ccaggatgaa
     1621 gtgccacagc tggatctgcc gggagcctgg ggatgactct gttttgtagg cccagtcagg
     1681 aaactgatac cctccacctc cttctggctt ttccacgatg ctacagccgg ctttcatatg
     1741 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
     1801 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
     1861 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
     1921 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     1981 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     2041 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     2101 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     2161 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
     2221 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     2281 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     2341 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     2401 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     2461 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     2521 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     2581 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     2641 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     2701 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2761 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2821 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2881 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2941 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     3001 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     3061 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     3121 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     3181 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     3241 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3301 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3361 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     3421 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     3481 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     3541 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     3601 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     3661 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     3721 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     3781 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     3841 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     3901 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3961 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     4021 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     4081 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     4141 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     4201 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     4261 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     4321 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4381 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4441 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     4501 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc
     4561 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     4621 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4681 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4741 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4801 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4861 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4921 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4981 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5041 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5101 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5161 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5221 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5281 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5401 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg
     5461 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     5521 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     5581 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     5641 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     5701 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     5761 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     5821 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     5881 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     5941 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     6001 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     6061 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     6121 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     6181 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     6241 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     6301 tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6361 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     6421 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     6481 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     6541 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6601 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6661 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6721 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6781 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6841 tacagggcgc gtcccattcg cca
//
