LOCUS       sequence-258596-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ETV4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  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 6753)
  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: ETV4_pet28a
FEATURES             Location/Qualifiers
     source          1..6753
                     /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(1631..1648)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1658..1675)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1694..1716
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1731..1755
                     /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(1755..1774)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1756..1774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1858..1877)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2083..2160
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2161..3243
                     /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(2180..2199)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4052..4243
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4237..4259)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4345..4484
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4400..4419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4499..4516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4670..5258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4750..4769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5369..5388)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5380..6195
                     /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             5380..6195
                     /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(5453..5472)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6287..6742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6424..6445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6636..6655
                     /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 gcctagtaag
      181 agtagccacc cttggggcca aatggctggg cggggccagc cagctctggg aggtaggcgg
      241 ggctctcatc caagtgggac aaagggactg tgtcctcctc actgacaggc cggtcaaact
      301 cagccttgag agctggacgc tgattgtccg ggaaggccaa agagaagagg gcctcgggct
      361 cacacacaaa cttgtacacg taacgctcac cagccacctt ctgcatgatg cctttctcat
      421 aatagtatcg gagcgagcgg ctcagcttgt cgtaattcat ggctggccgg ttcttctgga
      481 tgccccagag cctggcgacc tcctcaggct caatgagctt gaactccatt ccccggcccg
      541 tccaggcaat gaaatgggca tttgttgggt catccagcaa ggccaccaga aattgccaca
      601 gctgcagggc accccggcgc tggtagggcg gcccctctcg aaatgcaccg accccttcct
      661 gcttgatgtc tccttcaaat ttctcaggga caacgcagac atcatctggg aatggtcgca
      721 gaggtttctc atagccatag cccatggccc cgtcacctgg agagggccca gagaagccct
      781 ctgtgtggag gtacattgat gcgcacccgg tgacatctga gtcgtaggcg aagtccgtct
      841 gttcctgttt gatcaccacc cccgcccctg ggtacctgtg cccattgacc ccaccctggt
      901 ccacggctgg ctggcccgcc tgttcataca ggggatcatg gtattcttgc ttaaagctct
      961 gctggggata gggtgggcag ggctccgaca gctggtgttg gtagggggct gggaggggtt
     1021 cccggccccc tccctgagat gtgaaggagt ggcaaatgtc caggggctgc tggaagacgg
     1081 agctatgttc cccgaggtac ccatggccag ggtggggctg ggaggtgcca gaggatctca
     1141 ggaaattccg ttgctctgcc cggggaaagg gctgtagggg cgactgtcca agggcaccag
     1201 gggcagggga cttgatggcg atttgtctgg gggggtcata ggcactggag taaaggcact
     1261 gctcgccatg gtggtagggg agtggcggct tcctgctgca ggacagggcc gggtctgtgc
     1321 ggggactctg gggctccttc ttgatcctgg tggtggggct gtggaaagct aggttttctg
     1381 aatggaaatc aggaacaaac tgctcatcac tgtctggtac ctgagcttca gcgagccacg
     1441 tctcctggaa gtgacttaga tcctggaaga gatcttcaga gtcgaggggc ggcagggagc
     1501 ccgggtccat gagcttcccc agcgggccga tcagcgcttc gcgcaagctc ccatttccgg
     1561 gcgatttgct gctgaaggtg tagggcactt gctggtccaa gtatccggct ttcatcctcc
     1621 gctccatatg gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc
     1681 tgcccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta
     1741 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
     1801 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     1861 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     1921 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     1981 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     2041 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
     2101 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     2161 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     2221 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     2281 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     2341 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     2401 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     2461 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     2521 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     2581 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     2641 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     2701 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     2761 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     2821 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     2881 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     2941 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     3001 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     3061 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     3121 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     3181 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3241 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat
     3301 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     3361 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     3421 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     3481 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     3541 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat
     3601 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     3661 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     3721 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     3781 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     3841 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     3901 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     3961 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     4021 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     4081 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     4141 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     4201 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4261 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4321 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4381 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt
     4441 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     4501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5101 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata
     5341 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     5401 acgtcttgct ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     5461 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     5521 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     5581 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     5641 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
     5701 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     5761 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     5821 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     5881 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     5941 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     6001 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     6061 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     6121 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     6181 gatgagtttt tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     6301 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     6361 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     6421 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     6481 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     6541 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     6601 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     6661 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6721 aatgcgccgc tacagggcgc gtcccattcg cca
//
