LOCUS       sequence-258712-        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ELF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  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 7292)
  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: ELF4_pet28a
FEATURES             Location/Qualifiers
     source          1..7292
                     /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(2170..2187)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2197..2214)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2233..2255
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2270..2294
                     /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(2294..2313)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2295..2313)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2397..2416)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2622..2699
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2700..3782
                     /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(2719..2738)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4591..4782
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4776..4798)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4884..5023
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4939..4958
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5038..5055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5209..5797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5289..5308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5908..5927)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5919..6734
                     /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             5919..6734
                     /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(5992..6011)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6826..7281)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6963..6984)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7175..7194
                     /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 gctattatat
      181 gtcatggggc tccatcttaa tgagggaagt agggttgaat ggggagaaag gggctggggt
      241 gggggatctt gtcagaaggc tcccagatgt ggtcacacta ggctcagcca tcagcaggga
      301 ccctgagcct gaggagagct caagttcagc cactggggtc agcccaacag tggggcggct
      361 gggaggagac aaagtctggt tgcccagaag gctcggattg tgggaacccc tggaaaccac
      421 ctgggttgga agtggctgaa gatgagcctg atctctcagg agctccctca gggtctcttc
      481 aggaaccagc agcccctcca tgggggcccc cgtcaccata ccctgaacat aggaggagga
      541 cctcagtggc ccctccttga ccctaggggc tgctgtagtg ccagaagtcc tgatgaaggc
      601 agcaatgaca gtcccagggg gctgagagct gggccctgct ggtccagccc ctgtgactgt
      661 gggtggcgcc gggttggtcg gacggttggc cccagccaga agttggggga ggccactgag
      721 aatcagtgga gcccctgggg ctgccacctg gctgtcttgg aaactggcgt tcagggggaa
      781 agaggcctgc aaagtgaagg tgtccttgaa agtagaggcc gctggaacac tctggagaac
      841 gagctgagtg ggagcagtag tactggcagg aggcccattg gtcagcaccg tggtcagtgg
      901 gatcgtctgc agggtcaggg ccgagcccga ccccacgggg gccactccta ggtggatgtt
      961 gctgggcact gaagatgcac tcacagcttt ggtgagcttg acctggccct ctgctggaga
     1021 gacgagcatg gtggaggtag tggggatctc ctcgtctagc gacggtccca attccagact
     1081 cgcagatggc tggagaccga catgctgaat ttttggcttc tcccaagaag agctgccctt
     1141 gccctggggg gcagatctgg atgagaccct ggagctggtt cgccgggtgg tactggcaga
     1201 ggccacagag gccgtggagg cctgaggtgg ggctgctgtg gcttcgctgc tctcatcctc
     1261 atcttcaatg accaccaggt ccttgggcat ctccttaaac tggtacacca gcctctgccc
     1321 ttccactttg gccagtatgc ctctttggta gtagtatctt agtgcccgcc ccattgtctc
     1381 atagttcatg tcaggcttgt ttttctgctt cccccacagc ttggacacag ctttggagtc
     1441 caccagtttg aagatgcctt tctctcgctg ggtccacttg atgtacttgg gacaggtgtt
     1501 tctgtcttgc agaagagcca ggaggaactc ccacagatag atggtgctgc ctttgccatc
     1561 ctttgatttc ttcctaatgg ggatgctggg gtcagtgaca ggtgaggtac ttcggttgcc
     1621 cttggtcttc cggattctct tctttgattt cccagtcttc ttggcacttt cctctctgga
     1681 ggccttctcc tccagagaat gcccctcctg gtcactagtg tcacccgccc tgttcagggc
     1741 atcgggctca gaggcaggaa acaggtagtt gggcagagtg acagctggtg cagggtccga
     1801 gtctggaagc atttcggagg tactgaagat ctgcttctca tccaggatat cgctgggaga
     1861 ctccatattg agtaagactt ccgcggttga catggtgtgc gaggtggcct cattgtcatc
     1921 tgtcagcaaa aaactgcctt ccaggatctg atcctgcgtc atgcacaagg tcccgtctgt
     1981 tatgatgcca ttgtgaacgt cgtccaactc cagtcccgag tacagatgca gtaaatcagg
     2041 gtagggtacc tgctccacga tcacagctgg gaacacagag gggtcttcca gctggtggat
     2101 atcatcatcc atcccgttgc ttgcgaactc aaagatcagg tcactgggct gtagggtaat
     2161 agccatatgg ctgccgcgcg gcaccaggcc gctgctgtga tgatgatgat gatggctgct
     2221 gcccatggta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
     2281 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct
     2341 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     2401 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     2461 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     2521 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     2581 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
     2641 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     2701 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     2761 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     2821 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     2881 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     2941 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     3001 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     3061 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     3121 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     3181 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     3241 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     3301 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     3361 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     3421 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     3481 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     3541 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     3601 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     3661 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     3721 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3781 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
     3841 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3901 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3961 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     4021 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     4081 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc
     4141 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     4201 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     4261 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
     4321 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     4381 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
     4441 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
     4501 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
     4561 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
     4621 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     4681 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     4741 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4801 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4861 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     4921 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg
     4981 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     5041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5641 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa
     5881 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     5941 cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     6001 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     6061 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     6121 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     6181 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     6241 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     6301 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     6361 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     6421 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     6481 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     6541 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     6601 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     6661 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     6721 atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6781 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     6841 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     6901 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     6961 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     7021 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     7081 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     7141 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     7201 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     7261 atgcgccgct acagggcgcg tcccattcgc ca
//
