LOCUS       sequence-258708-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ELF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  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 6875)
  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: ELF2_pet28a
FEATURES             Location/Qualifiers
     source          1..6875
                     /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(1753..1770)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1780..1797)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1816..1838
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1853..1877
                     /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(1877..1896)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1878..1896)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1980..1999)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2205..2282
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2283..3365
                     /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(2302..2321)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4174..4365
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4359..4381)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4467..4606
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4522..4541
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4621..4638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4792..5380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4872..4891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5491..5510)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5502..6317
                     /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             5502..6317
                     /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(5575..5594)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6409..6864)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6546..6567)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6758..6777
                     /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 gctattattt
      181 ctcacatgtc actagtcctt ctgttttcat agttacagga agggcaatag ctgaaggcgc
      241 actgacaacc actacgtggg tcactgtctt atttccatct gctggttttt cttctactag
      301 ctgcaaagtt ttcacatcat gttcttgctt ttttgccact gcttccgatt taacctctgg
      361 cccctttatg actgcactga taactcgagg aggagtctgg ccagatgcct gctgagtagg
      421 cattgatagt ctcattacag gtgtaccatg ggctattgaa acaggggtaa gtgctctcac
      481 agccaatggg gttccaacaa tgttaatgct tcctgatcca gtcagatttg actttgtctg
      541 cagttgacac tgtgcaagct gtgtagctgg gatggtaata attttggcag gctgcatggt
      601 gattttgtct ccattttcag tagaagctgg catcacagta gggattgtct gaatgactac
      661 ctttggagag gtcgctgttg ttggactagt gctggttatt aatggtgcac ctgcattaac
      721 tgactgaact gccacagttg aaattttctg acccaatgat gtcattacaa caggtacctg
      781 cattgccaca cgaactgtcc ttggagctgc tgttgctgac acagatgcag tggtagtagg
      841 agacctggat gaagcatcgt gcccagggga agtgatattc acaactctag ctacaccctt
      901 ctctgctctg gagcagttta taggggatga attttttcca ctgcgaacag aggatgctgc
      961 tttcaggaga ctttctgcag acagtgacac tcgttctaat gatttttcat cagtagttcc
     1021 tgctaaatct tcattacagg tttcactttt gtcatcatct atgaccacta tgtttttcgg
     1081 catatccttg aactgatata caagcctctg tccttcaacc tttgcaagaa ttcccctttg
     1141 gtagtagtat ctcaaagctc gtcccatggt ttcatagttc atgtctggtt tgttcttatg
     1201 ctttccccaa agcttagaga cagcctttga atccaccagc ttgaatatgc ctttttctct
     1261 ctgagtccat ttaatatacc tgggacaagt atttttatct tgaagtagat ctaaaagaaa
     1321 ctcccacaaa taggttgtgt ttccttttcc ttctcttggt ttcttcttta tacctaactc
     1381 aggagaccca ttggaaattg gtgattgctg ggtctttggt ttacggccaa cttttttctt
     1441 tttcattggt tcatggctat ctggtgatgt tggaatagga gaggtatcca tgggttcaga
     1501 ctcttcagtt gacacctcca ctacagtttc tgtaatgaca tctggcctca tagcagcatg
     1561 gatgaattca ggacttcttg aatccctcaa gcaggtagga gattccatat gaagcagggc
     1621 ttcagcagct tcaattgtct tatctgtaca gtgtgcatta ctgctgtgaa ctgatgcttc
     1681 cacggcccgg atgagcagat ccagctggtt cgtgggtccc tcatgcagag acgtcgccat
     1741 gctagccata tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct
     1801 gctgcccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
     1861 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat
     1921 cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc
     1981 ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc
     2041 ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc
     2101 cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg
     2161 cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc
     2221 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     2281 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     2341 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     2401 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     2461 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     2521 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     2581 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     2641 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     2701 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     2761 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     2821 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     2881 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     2941 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     3001 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     3061 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     3121 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     3181 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     3241 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     3301 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3361 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg
     3421 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     3481 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     3541 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     3601 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     3661 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg
     3721 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     3781 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     3841 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     3901 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     3961 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     4021 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     4081 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     4141 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     4201 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     4261 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     4321 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4381 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4441 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     4501 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg
     4561 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     4621 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4681 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5221 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa
     5461 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     5521 aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     5581 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     5641 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     5701 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     5761 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat
     5821 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     5881 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     5941 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     6001 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     6061 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     6121 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     6181 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     6241 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     6301 tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa
     6361 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     6421 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6481 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6541 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6601 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6661 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6721 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6781 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6841 ttaatgcgcc gctacagggc gcgtcccatt cgcca
//
