LOCUS       sequence-258526-        6741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ETV1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6741)
  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 6741)
  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: ETV1_pet28a
FEATURES             Location/Qualifiers
     source          1..6741
                     /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(1619..1636)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1646..1663)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1682..1704
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1719..1743
                     /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(1743..1762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1744..1762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1846..1865)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2071..2148
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2149..3231
                     /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(2168..2187)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4040..4231
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4225..4247)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4333..4472
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4388..4407
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4487..4504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4658..5246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4738..4757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5357..5376)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5368..6183
                     /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             5368..6183
                     /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(5441..5460)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6275..6730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6412..6433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6624..6643
                     /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 gcttaataca
      181 cgtagccttc gttgtagggg tgggggttgc agcagccccc ttccggcatg taggccatgc
      241 tctcatcaaa gtgagaaaga ggcactgtgt cctcctcgtt gatgtgacgt tccatgtctg
      301 tcttcagcag tggacgctga ttatctggaa aggccatgga gaaaagggct tctggatcac
      361 acacaaactt gtagacatat ctctctccag ccaccttttg cataattcct ttctcatagt
      421 aatagcggag tgaacggcta agtttatcat agttcatagc tggcctgttt ttctgaatgc
      481 cccaacgtcg ggccacctct tcaggctcaa tcagtttaaa ttccatgcct cgaccagtcc
      541 aggcaataaa atgagaattt gaagggtcat ccagaagagc taccaaaaac tgccagagct
      601 gaagtgatcc tcgccgttgg tatgtgggtc cttcccgata cattcctggc tcttgtttga
      661 tgtctccatc gaatttttct gggacaacac aggtgtcatc ataaaactgc ctggggccct
      721 tttcaaacat acagccttct gttctgctgg gatgagccag gaagccttct tgcctcatat
      781 aaatggagtg gcagctaggc acttctgagt catatgcaaa atctctgggt tcctgtttaa
      841 tcatcagagg agggggaaag ctttggctgg ccgcactgcc aaccatggtg ttgtgttcat
      901 acactgggtc gtggtactcc tgcttaaagc cttgtggtgg gaaggggatg tttggctcag
      961 acatctggcg ttggtacata ggacgtcctt cccttggcat cgtcggcaaa ggaggaaagg
     1021 agttacaggg ttcagaaagc tggcggcgaa atctgtggtc catggggtag ctgctatctg
     1081 gtatggactg cgatggaggg aggtgagctg ggaaggcccg gtcaggtttc ggtgtatgag
     1141 ttgagtttgg agatgcatga tgcagtgggg acactggcgt gctggatggt gtgggggggt
     1201 tggagggcct cattcccact tgtggcttct gatcataggc actgacattg tacaggcact
     1261 tttctccata gctgaattta aagggctgtt cttgactgca ggcagagctg atttctgaac
     1321 atggactgtg gggttctttc ttgattttca gtggcaggcc atgaaaagcc aaactttcag
     1381 cctgatagtc tggtacaaac tgctcatcat tgtcaggtac ctgagcttct gcaagccatg
     1441 tttcctgtaa ttgacttaga tcttgaaaga gttcttctga atcatgagcc agatctctgt
     1501 taatgaattt tcttttcctg acatttgttg gtttctcgtt acaatttctc ccacgctgac
     1561 tattggtgac catgtaaggc acttgctggt cataaaatcc atccatgcta gccatatggc
     1621 tgccgcgcgg caccaggccg ctgctgtgat gatgatgatg atggctgctg cccatggtat
     1681 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat
     1741 tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac
     1801 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     1861 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     1921 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
     1981 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
     2041 agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc
     2101 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     2161 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     2221 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     2281 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     2341 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     2401 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     2461 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     2521 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     2581 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     2641 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     2701 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     2761 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     2821 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     2881 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     2941 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     3001 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     3061 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     3121 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3181 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3241 aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc
     3301 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     3361 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     3421 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     3481 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     3541 gaagcaggcc attatcgccg gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc
     3601 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
     3661 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
     3721 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
     3781 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     3841 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
     3901 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
     3961 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
     4021 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     4081 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     4141 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     4201 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4261 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4321 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     4381 aactatgcgg catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc
     4441 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4501 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4561 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4621 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4681 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4741 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4801 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4861 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4921 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4981 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5041 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5101 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5161 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5221 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5281 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
     5341 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
     5401 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     5461 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     5521 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     5581 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     5641 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     5701 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     5761 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     5821 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     5881 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     5941 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     6001 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     6061 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     6121 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     6181 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6241 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     6301 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6361 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     6421 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6481 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     6541 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     6601 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     6661 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6721 cagggcgcgt cccattcgcc a
//
