LOCUS       sequence-258702-        6410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene ETV3L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6410)
  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 6410)
  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: ETV3L_pet28a
FEATURES             Location/Qualifiers
     source          1..6410
                     /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(1288..1305)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1315..1332)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1351..1373
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1388..1412
                     /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(1412..1431)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1413..1431)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1515..1534)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1740..1817
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1818..2900
                     /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(1837..1856)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3709..3900
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3894..3916)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4002..4141
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4057..4076
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4156..4173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4327..4915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4407..4426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5026..5045)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5037..5852
                     /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             5037..5852
                     /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(5110..5129)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5944..6399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6081..6102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6293..6312
                     /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 cgaattcatt aaggaggatc caagggccac actgctttga ggttctccag
      241 attgggggaa gtaaggcttt cctccccagt tttgagtctg cgggtctctg ggcagaagac
      301 ctccctggga tccaggcctc cctttgcctc catcatggga gcaggcttta cttccagccc
      361 ctcgggcctg agggacaaga gccaaagcct ctcacccgca ccctgtccca gccctgccaa
      421 gagaggaagc cctggaaaat gccaggcccc tgggaggctc ctaggtcctg ggagcaggat
      481 gtctggctta aaagcccctg ggagctgctg ctctgacggg agaggaggca agaagggccc
      541 cgagagacag gtccagttgg agggcagagg gggcgggagg ggaggagtga aggaggcaac
      601 accaggcagc tcgccttgga cgctccccaa atggcagcaa aggcccagcc ggcaggggcc
      661 tggggcagag ccaagtcggt agacgctgct gctgctcccc ttcttatccc cagaggtctc
      721 tggtggccct cggggggtct gctgtcctgt cagctgctcc accatggcct gatgggcaaa
      781 tagcatgctg tgcaggagct cactctgcac acccacgggc accagcgctg gccgacacag
      841 ggcaggggcc cccagcagca agtgggggga tggcggcgcc cgcacttccc acaaaggata
      901 gttgactacg atgagcttgc tgaagttgaa cttgtatgtg aaccttttgc ctttggtctt
      961 atgcaggatc ctcttattgt agtaatatct gagggcccgg ctcagcttgt cataattcat
     1021 ctgtggtttg catttcctgc ggccccagag gcgggccacc tcatctggat ccttgatgac
     1081 aaattccccg tactctccct gctgccaggc gatgacatgg cggaactctt ccttctgcag
     1141 cagctccagg atgaagtgcc acagctggat ctgccgggag cctggggacg actcggcttt
     1201 gtaggcccaa tcagggaagg ccaaccctga gatccagttg ccggggttgg ctgggatgcc
     1261 ctcagccaag cagctgcagt gcatatggct gccgcgcggc accaggccgc tgctgtgatg
     1321 atgatgatga tggctgctgc ccatggtata tctccttctt aaagttaaac aaaattattt
     1381 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
     1441 gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     1501 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     1561 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     1621 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     1681 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg
     1741 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     1801 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     1861 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     1921 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     1981 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     2041 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     2101 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     2161 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     2221 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     2281 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     2341 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     2401 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     2461 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     2521 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     2581 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     2641 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     2701 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     2761 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     2821 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2881 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     2941 ccgggatctc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     3001 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     3061 caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
     3121 atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
     3181 actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc
     3241 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
     3301 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
     3361 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
     3421 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
     3481 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga
     3541 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc
     3601 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc
     3661 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag
     3721 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
     3781 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
     3841 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     3901 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3961 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     4021 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     4081 gagtgcacca ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4981 tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     5041 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
     5101 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     5161 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     5221 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     5281 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     5341 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     5401 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     5461 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     5521 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     5581 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     5641 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     5701 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     5761 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     5821 agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt
     5881 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5941 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6001 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6061 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6121 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6181 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6241 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6301 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6361 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
