LOCUS       sequence-258531-        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ELK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  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 6616)
  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: ELK1_pet28a
FEATURES             Location/Qualifiers
     source          1..6616
                     /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(1494..1511)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1521..1538)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1557..1579
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1594..1618
                     /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(1618..1637)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1619..1637)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1721..1740)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1946..2023
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2024..3106
                     /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(2043..2062)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3915..4106
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4100..4122)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4208..4347
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4263..4282
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4362..4379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4533..5121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4613..4632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5232..5251)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5243..6058
                     /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             5243..6058
                     /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(5316..5335)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6150..6605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6287..6308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6499..6518
                     /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 cgaattcgga tcctcatggc ttctggggcc ctggggagag caccacgggg
      241 gtcgagaggc catccacgct gatagaaggg atgtgcacct gggcgctgcc actggatgga
      301 aactggaagg agagcttggc cgggctacgg ggcgcaatgg gactcagggt gctccagaag
      361 tgaatgctag gaggcagcga gctgggtgtc agcagcaccg gggtcaatgt atgcgtagga
      421 agcagggatg gggtcagcgc cggccccgga gcctggaggc cggagccact tcccgatcct
      481 ggggtccgtt cgggtcccgg cccacctagc aggctcgggc tgagtggaag ctctaggtcc
      541 cggggcttcc ggcccttctg cggctgggag atctcagggc tggaagccgc atggccgccc
      601 gcctgccctg cggtgtccat aacaaccgcg ggcagccggg ctggcacgcc ctcctgtgga
      661 gggacttctg gctcggcctt ggtggtttct ggcacaaacc ctctctcccc cgcaacttcc
      721 aactcttcct tgggcccttc tactttcact tctgggggca aagcccggcc caaacccggc
      781 tccacattaa gctcttccga tttcaggttt ggggcctcgg gcggggtcag gatgacctgc
      841 agaggcaagc cggcctcttc agcctccaga caggcctcca aggggcttgg actggtgctc
      901 ctgctccccg aggggggcgc tgctgcccct gcaggagctg cattggggag caccacagca
      961 ggccgaggat gagggggtgg ctgcggctgc agagactgga tggtgaaggt ggaatagagg
     1021 cccgagcgca tgtactcgtt ccggctgctg cgtgccaaac cgcctgggcc tgccattcct
     1081 gcacccttgg gtgtgcctgg ctttccagag acagtgtccc ctggggcggc atgtatagca
     1141 gcaggggcca catttggcat ggtggaggta acagacacct ctggctgggg cgggcagtcc
     1201 tcagtggagc accctgcgac ctcagggtag gacacaaact tgtagacgaa cttctggccg
     1261 ctcaccttgc ggatgatgtt cttgtcatag tagtaccgca aggcccggct gagcttgtcg
     1321 taattcatgt tggtcttgtt cttgcgtagt ccccacagcc gggccacctc ctctgcatcc
     1381 accagcttga attcaccacc atcccgtgaa gtccaggaga tgatgtggcc attgccttgc
     1441 tctctcagca gctgcagcag aaactgccac agcgtcacag atgggtccat atggctgccg
     1501 cgcggcacca ggccgctgct gtgatgatga tgatgatggc tgctgcccat ggtatatctc
     1561 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
     1621 tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc
     1681 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     1741 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     1801 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     1861 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     1921 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
     1981 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     2041 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     2101 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     2161 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     2221 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     2281 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     2341 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     2401 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     2461 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     2521 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     2581 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     2641 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     2701 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     2761 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     2821 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     2881 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     2941 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     3001 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3061 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3121 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
     3181 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     3241 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     3301 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     3361 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     3421 aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga
     3481 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     3541 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     3601 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     3661 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     3721 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     3781 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     3841 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     3901 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     3961 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     4021 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     4081 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4141 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4201 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     4261 tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga
     4321 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     4381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5161 tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt ctgcttacat
     5221 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctctaggcc
     5281 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     5341 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     5401 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     5461 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     5521 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     5581 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     5641 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     5701 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     5761 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     5821 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     5881 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     5941 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     6001 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaga
     6061 attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6121 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     6181 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6241 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6301 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6361 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6421 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6481 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6541 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6601 cgcgtcccat tcgcca
//
