LOCUS       sequence-258589-        6619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ELK4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  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 6619)
  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: ELK4_pet28a
FEATURES             Location/Qualifiers
     source          1..6619
                     /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(1497..1514)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1524..1541)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1560..1582
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1597..1621
                     /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(1621..1640)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1622..1640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1724..1743)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1949..2026
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2027..3109
                     /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(2046..2065)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3918..4109
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4103..4125)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4211..4350
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4266..4285
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4365..4382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4536..5124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4616..4635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5235..5254)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5246..6061
                     /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             5246..6061
                     /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(5319..5338)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6153..6608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6290..6311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6502..6521
                     /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 cgaattctta tgtcttctgt aggtctgggg aaaatgggcc aggggtggaa
      241 ggtccatcca gcccagacag agtgaatggc ccatgactgt tcagtacaga aggaaactgg
      301 aaaagtgtgt tagcaccttg cagtctggct ggacttaggg gagcaacagg actgagagta
      361 ctccagaagt ggatactgga gagcaagggg cttggagtca gtatgatggg tgtctgtgaa
      421 aaaaatgctg gtgtaagaga agctgtaggg agagatgggc tcagtattcc cagtgggctt
      481 ggatcactgc tcgtgatcac aagggtgggt gccagttcta accctttggg tttcttggat
      541 cttgatgaat tatttacttt gtccttttct agcaagactg aatcctggtc tttaggctcc
      601 agtgacaaat tctctggaag ttccattggc tgagaagcca ctgaatcaat gtctgtgtcg
      661 atgtctgggt gagaactcag tggtggtgaa ggtgttctgg gaggttcctg caaagggggt
      721 atggacgaaa tgggtggtgt ggtggcaaaa gcagtcatta cgttagaggc agaggttggg
      781 gcttccaggg aaggcagttt tggggaaacc aatgtctcca aagcttggat agtttcttct
      841 gaagatggag aaatacttgg gccaattgaa atggtggcag caacaggttc aaccggtggc
      901 tttttggaag gtgtcgtgac aaatttgatg acagatggtg tgggctcctg aggagatttt
      961 ttctctgcca gtttctcggc tggattctca gtctttatca atttgaaaag ctttacattg
     1021 gaggagttca aagagttgag agtaaatgaa gaatataagc cagagtgtat gtagtcattg
     1081 cggctagagg tcttggcacc aggctgaggt ggtttatctt tccctccatt ctccacatct
     1141 ttggaactgc tgctgacttc actgaagttt aaactttcac agtcaccctc aatcctgccc
     1201 actgtcattg gatccatgtt caaaatctct ggataagaga caaacttgta cacaaacttc
     1261 tgaccattca cttttttgat gatattcttt acataatagt atctgagggc tcggctgagt
     1321 ttgtcataat tcatgttagg cttgttcttg cgaatccccc agagacgagc cacctcttct
     1381 gcctgcaaaa gcttaaactg cccatcatta gaggtccaac agatcatgtg cttgttctga
     1441 ggcttctgca ggagctgaag aaggaactgc cacagggtga tagcactgtc catatggctg
     1501 ccgcgcggca ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat
     1561 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
     1621 ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc
     1681 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     1741 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     1801 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     1861 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     1921 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
     1981 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     2041 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     2101 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     2161 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     2221 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     2281 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     2341 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     2401 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     2461 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     2521 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     2581 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     2641 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     2701 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     2761 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     2821 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     2881 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     2941 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     3001 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3061 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3121 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     3181 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     3241 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     3301 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     3361 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     3421 agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt
     3481 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     3541 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     3601 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     3661 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     3721 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     3781 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     3841 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     3901 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     3961 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     4021 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     4081 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4141 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4201 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     4261 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac
     4321 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     4381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta
     5221 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag
     5281 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     5341 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     5401 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     5461 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     5521 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
     5581 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     5641 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     5701 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     5761 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     5821 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     5881 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     5941 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     6001 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     6061 agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6121 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6181 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6241 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6301 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6361 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6421 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6481 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6541 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6601 gggcgcgtcc cattcgcca
//
