LOCUS       sequence-258593-        6747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ERG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6747)
  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 6747)
  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: ERG_pet28a
FEATURES             Location/Qualifiers
     source          1..6747
                     /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(1625..1642)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1652..1669)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1688..1710
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1725..1749
                     /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(1749..1768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1750..1768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1852..1871)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2077..2154
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2155..3237
                     /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(2174..2193)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4046..4237
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4231..4253)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4339..4478
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4394..4413
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4493..4510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4664..5252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4744..4763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5363..5382)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5374..6189
                     /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             5374..6189
                     /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(5447..5466)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6281..6736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6418..6439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6630..6649
                     /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 gcttagtagt
      181 aagtgcccag atgagaaggc atatggctgg tggggagcct agtgttgggg tatatacccc
      241 cagttggtga attccagtat gggtttgggg cagcaaaaaa actggaagat gtcacgggga
      301 gggctggagg gtggggcgcc acaaagttca tcttctgtgg gtgggcgtga taggagccca
      361 tgtacgggag gtctgagggg tacttgtaca gagatgactc cggggggtgg ggctggaggg
      421 cctgggcgat cccgtggaag tcgaacttgt aggcgtagcg cttcccatgg accttggtca
      481 tgatgttctt gtcatagtag taacggaggg cgcggctgag cttatcgtag ttcatgttgg
      541 gtttgctctt ccgctctccc cagcgccggg ccacctcgtc gggatccgtc atcttgaact
      601 ccccgttggt gccttcccag gtgatgcagc tggagttgga gctgtccgac aggagctcca
      661 ggaggaactg ccaaagctgg atctggccac tgcctggatt tgcaaggcgg ctacttgttg
      721 gtccaagaat ctgataagga tctaactgag gacgctggtc ttcagttttg ggcactgtgg
      781 aaggagatgg ttgagcagct ttcgactggg gcgtggggtg gccgtgaccg gtccaggctg
      841 atctcctggg gggctcatat ggtaaatctg gcctagttgt aattctttgc gtagcttcag
      901 gatatactga agtatttggg aaaataaaag ctgcaccccc tgtgtttcta gcatgcatta
      961 accgtggaga gttttgtaag gctttatcaa catcatctga agtcaaatgt ggaagaggag
     1021 tctctctgag gtagtggaga tgtgagagaa ggatgtcggc gttgtagctg ggggtgagcc
     1081 tctggaagtc gtccttggtc atcttgcaca gttccttccc atcgatgttc tggaataaca
     1141 agatgttgac gtctggaagg ccatattctt tcaccgccca ctccagccac tgccgcacat
     1201 ggtctgtact ccatagcgta ggatctgctg gcacgataac tctgcgctcg ttcgtggtca
     1261 tgtttggggg tggcatgtgc ttctcctcca tgtagctgcc gtagttcatc ccaacggtgt
     1321 ctgggctgcc caccatcttc ccgcctttgg ccacactgca ttcatcagga gagttccttg
     1381 agccattcac ctggctaggg ttacattcca ttttgatggt gaccctggct gggggttgag
     1441 acagccaatc ctgctgaggg acgcgtgggc tcatcttgga agtctgtcca tagtcgctgg
     1501 aggaggacgc ggtcatctct gtcttagcca ggtgtggcgt tccgtaggca cactcaaaca
     1561 acgactggtc ctcactcaca actgataagg cttccttaat agtgctggcc atgctagcca
     1621 tatggctgcc gcgcggcacc aggccgctgc tgtgatgatg atgatgatgg ctgctgccca
     1681 tggtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct
     1741 cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg
     1801 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     1861 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     1921 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     1981 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     2041 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     2101 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     2161 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     2221 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     2281 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     2341 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     2401 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     2461 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     2521 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     2581 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     2641 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     2701 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     2761 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     2821 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     2881 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     2941 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     3001 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     3061 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     3121 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3181 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3241 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     3301 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     3361 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     3421 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     3481 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     3541 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
     3601 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     3661 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     3721 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     3781 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     3841 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     3901 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     3961 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     4021 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     4081 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     4141 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     4201 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4261 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4321 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     4381 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg cggtgtgaaa
     4441 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     4501 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4561 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4621 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4681 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4741 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4801 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4861 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4921 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4981 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5041 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5101 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5161 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5221 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5281 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg
     5341 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     5401 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     5461 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     5521 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     5581 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     5641 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
     5701 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     5761 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     5821 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     5881 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     5941 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     6001 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     6061 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     6121 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     6181 tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     6301 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     6361 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     6421 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     6481 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     6541 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     6601 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6661 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6721 ccgctacagg gcgcgtccca ttcgcca
//
