LOCUS       sequence-258598-        7160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ELF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7160)
  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 7160)
  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: ELF1_pet28a
FEATURES             Location/Qualifiers
     source          1..7160
                     /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(2038..2055)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2065..2082)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2101..2123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2138..2162
                     /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(2162..2181)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2163..2181)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2265..2284)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2490..2567
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2568..3650
                     /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(2587..2606)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4459..4650
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4644..4666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4752..4891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4807..4826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4906..4923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5077..5665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5157..5176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5776..5795)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5787..6602
                     /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             5787..6602
                     /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(5860..5879)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6694..7149)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6831..6852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7043..7062
                     /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 gctactaaaa
      181 agagttgggt tccagcagtt cgttttgttt catagctacc tgagaagtaa atccattgga
      241 actggacact accatcacat aaggctgtgg ctgctgctcc gttttctcag tgttctcttt
      301 caaatgatct tcagattcct ttttctctac ttcctgtgta agagtttttg tttcttgagt
      361 tttgataact gaagtgatta cagtgccagg tgggtgagca accagctgtg aactgcgagg
      421 agaaaaggtc accacaggtg cagtagcact gaaggatgga gaggaagcca cactgacggt
      481 tccattgcaa atggtctgaa cattgctagt aaggacctgc tgaacctggg cagggcccaa
      541 gacaattgaa ggaggagagc ccgccttttg tgactgcagc atgacatttt ctttcagtac
      601 tgtcatgggc tgtgatgatg gaatggcttg taaaataaac ttctgagatc cagtacctgc
      661 tgatggatct gtgctggcta taactgttgt gagtggcact gtttggagtg ttactgtatg
      721 caactgctga ttcctaggag acacaaccac tggaacttgg gttggagcct gtatagtcct
      781 aatactctga acggaagaat ttaatgtttc atcctgcatg gtactggttc tagctgcttc
      841 tccctctggg acagcctgta ctggctgtac tacatgaaca gtccggaaga gctgggtagg
      901 atatggagac tgcgtgggct gcactgtcct caaaacttct gatggttgtg caacttccac
      961 aggatctttg ggttttgcag ctttagaatt ccctggtttt agaactgaag tggctcctcc
     1021 ttttacccct ggacttgaag atactctcga ccggctggtt tgattcctat ttgaagtggc
     1081 tgatgaagat agcgatggat ctgaagactc tatgctggaa cttggatcct catcatttat
     1141 atatataaga tcttttggca tttctttaaa ctgatacacc aagcgctgac cttccacttt
     1201 tgccagaata cccctttggt aatagtacct gagtgctctt cccatggtct cataattcat
     1261 atcaggtttg tttttgtgct tcccccacaa cctggacact gctttagaat ccaccaattt
     1321 aaaaatgcct ttctctcgct gggtccactt gatgtattta ggacaagtag ccttgtcctg
     1381 gagcagtgcc agtaaaaact cccaaagata aattgtgttt ccctttccat ctttgttttt
     1441 cttcttcaca gatatatttg gcgtagtggc tggggaatct ggtcgtggtg gtttagtttt
     1501 tcttcctttt ttcctcttag gctgttctgg tgatgaggct cccggtgagt ctgcatattt
     1561 ttcttgcacc tgctgtgttt ccatcacttc aggaatccca tctaatgtga cggacacatg
     1621 ggtgactggg gcaacaacca tgtcatcttc aggtgaacta aatatattat tatttattcg
     1681 tttttcatcc agcatagggc caggggaatc catattgagg agtgcctcag cagcctcaat
     1741 agtttcaatt gtttcatccc cgtcatgaca agaagcttca actgtaaggg tgatgtcatc
     1801 atcatcatcg tctatgattt cttcttcagc aacatccagt gaactctcag taatcatgtc
     1861 attgggctct tccacacagg ctagaccggc ataactattg agaatatcag caccaggaac
     1921 atgttccaca attacggcag gaaaaatagc tggatcacca agctgtcgtt catcctccat
     1981 gacgttacta gcaaattcaa atactaggtc gttctgttgg acaacagcag ccatatggct
     2041 gccgcgcggc accaggccgc tgctgtgatg atgatgatga tggctgctgc ccatggtata
     2101 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
     2161 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg
     2221 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     2281 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     2341 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     2401 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
     2461 gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg
     2521 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     2581 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     2641 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     2701 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     2761 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     2821 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     2881 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     2941 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     3001 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     3061 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     3121 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     3181 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     3241 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     3301 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     3361 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     3421 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     3481 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     3541 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3601 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3661 attaatgtaa gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc
     3721 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3781 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     3841 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     3901 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     3961 aagcaggcca ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg
     4021 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     4081 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     4141 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     4201 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     4261 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     4321 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     4381 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     4441 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     4501 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4561 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     4621 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4681 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4741 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4801 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
     4861 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     4921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5701 tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt
     5761 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta
     5821 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     5881 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
     5941 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     6001 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     6061 atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag
     6121 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     6181 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     6241 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     6301 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     6361 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     6421 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     6481 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     6541 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     6601 aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6661 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     6721 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6781 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6841 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6901 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6961 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7021 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7081 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7141 agggcgcgtc ccattcgcca
//
