LOCUS       sequence-258533-        6353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ETV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6353)
  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 6353)
  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: ETV2_pet28a
FEATURES             Location/Qualifiers
     source          1..6353
                     /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(1231..1248)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1258..1275)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1294..1316
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1331..1355
                     /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(1355..1374)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1356..1374)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1458..1477)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1683..1760
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1761..2843
                     /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"
     CDS             3652..3843
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3837..3859)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3945..4084
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4000..4019
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4099..4116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4270..4858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4350..4369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4969..4988)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4980..5795
                     /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             4980..5795
                     /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(5053..5072)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5887..6342)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6024..6045)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6236..6255
                     /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 cgaattcatt attgtgtctc tgctccccgt ccgcctcccg cacagtccgg
      241 ataggctagg ctgggcacgc ggcccccgaa gcggtacgtg tactttcgcc ccccgctctt
      301 gcgcacgatg tcgcggcgat agtagtagcg aaggccccgg ctcagcttct cgtaattcat
      361 gcccggcttt ctcttgcgct cgccccacag ccgagccacc tctttggggt cgcacagctg
      421 gaactcgcgg ctgttgccag tccaacggat gcagctgcta cgcgccccgt cgtggagcag
      481 ctccaggagg aactgccaca gctgaatggg acctcggtgg ttagttttgg ggcatcgagc
      541 caaactggca cggtccgact gcgggctcgg ttcggagcaa acggtgagag ctgagctctg
      601 gtaccgcttc aaagaggtgg tgccacccgc atgcagcccc gggttccagg aggtggtaca
      661 gtccgggccc gcgggcccgc cccacgaaat ggtacagtcc gtgcgcggct ccccgcccag
      721 gccactgccc cagtaggtat cgccgtcggg ccccacagaa cagtcccagc tggtgttgct
      781 cccggcggcc tgggcgcgcg accacgaggt ggcctctccc gccacgggga cgcagttctg
      841 gcccgcggcg ccttcggagc cggcggcggg gatggggccc gggccgagag gggcggggcc
      901 cagggtctgc gaggcgccgc tccaagagtc ccaggctgtg cacgccatgt ctgtccagtc
      961 cccggaccac ggaagagcct gagagtcggg ctccgccccc catggaactt ctgggtgcag
     1021 taacgcggag ccccagtcca gctgtgggaa gcttgccagg gatgagcttg tacctttcca
     1081 gcatgtctct gctgtcgctg tcggcgtgtc cccttggagt gccagatcag ggaaacagaa
     1141 gcctaatttg gctccttcaa gccctgccag cttgttccct ggaggcactt cctgtgggga
     1201 tgcctcatcc cagttccaca ggtccatatg gctgccgcgc ggcaccaggc cgctgctgtg
     1261 atgatgatga tgatggctgc tgcccatggt atatctcctt cttaaagtta aacaaaatta
     1321 tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
     1381 cgggatcgag atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     1441 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     1501 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     1561 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     1621 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc
     1681 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     1741 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     1801 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     1861 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     1921 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     1981 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     2041 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     2101 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     2161 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     2221 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     2281 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     2341 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     2401 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     2461 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     2521 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     2581 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     2641 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     2701 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     2761 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2821 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     2881 gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     2941 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     3001 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     3061 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     3121 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     3181 gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     3241 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
     3301 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
     3361 agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga
     3421 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc
     3481 tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt
     3541 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
     3601 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
     3661 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
     3721 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
     3781 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3841 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3901 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     3961 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4021 tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     4081 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4141 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4681 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4921 gattttggtc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     4981 tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc aacatggatg
     5041 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     5101 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     5161 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     5221 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     5281 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     5341 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     5401 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     5461 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     5521 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     5581 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     5641 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     5701 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     5761 tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5941 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     6001 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     6061 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6121 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6181 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6241 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6301 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//
