LOCUS       sequence-258717-        6439 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression of ETS gene ELF3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6439)
  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 6439)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: ELF3_pet28a
FEATURES             Location/Qualifiers
     source          1..6439
                     /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(1317..1334)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1344..1361)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1380..1402
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1417..1441
                     /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(1441..1460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1442..1460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1544..1563)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1769..1846
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1847..2929
                     /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(1866..1885)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3738..3929
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3923..3945)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4031..4170
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4086..4105
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4185..4202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4356..4944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4436..4455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5055..5074)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5066..5881
                     /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             5066..5881
                     /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(5139..5158)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5973..6428)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6110..6131)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6322..6341
                     /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 cgaattctca gttccgactc tggagaacct cttcctcctt ccagccgctt
      241 gagtttttgc caaacttgta gacgagtcgc cggccatcca cccgttccag gatctcccgt
      301 ttgtagtagt acctcatggc ccggctcagc ttctcgtagg tcatgttgct gttctttttc
      361 ttttggcccc atagttgggc cacagcctcg gagcgcagga acttgaagac gccttcatgc
      421 cgattctccc acttcatgag gccctcgttg agctccgggt ggatgaggat gtcccggatg
      481 aactcccaca ggtgggtgcc tctgggcgcg tgcttgctct tcttgccctc gagacagtcc
      541 cagtactctt tgctcagctt tcggggccgg cctcgtttcc gcttcccgtg cttgggatcc
      601 cccttcttgc agtcacgaaa accatcgctg gggaagagct tgccatcagt gggatccagg
      661 tccacgtcac ttccaccgga gtctgaggag tgggagctcc gagaagcacc agtccctgcg
      721 gtggagacgt cagagctgcc aggggagggg gctcctgcgc cacagctgcc ggggtggtag
      781 gggctggctt gctgaccgtc gtccagcagc tcctgggcaa aggggctgcc ctggtcaaag
      841 ggccctgggt ctagggcctc ctggaaggcc atgccatcct tctccagcag ctcaatgatc
      901 caactgagct catcagaaga gctggaagtg aggtctcgca gctgggcatg gagttggtcc
      961 cccagaggcc caaagaccag acgcagctcc tcaagggcac aattgcagag ggtggcgcca
     1021 tccatgtcac atcgtgagaa gtcaatggcg cttgcgtcgt acttgttctt ctccacttgg
     1081 tagctgatcc agtccagaac ctgcgtcttc gaccagaact ggggctgttc ccccaaccag
     1141 ctggccttct ctgtaccctc caatgacatc tgggggttgc tcagggtcag taccaagtca
     1201 tcggccccaa aggtggcagc agggggaaca gaggccaggg tggagtcctc cgagctgtac
     1261 atcgcactga agtagttgct aaaaatgttg ctaatctcac aggttgcagc catatggctg
     1321 ccgcgcggca ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat
     1381 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
     1441 ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc
     1501 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     1561 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     1621 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     1681 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     1741 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
     1801 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     1861 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     1921 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     1981 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     2041 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     2101 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     2161 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     2221 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     2281 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     2341 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     2401 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     2461 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     2521 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     2581 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     2641 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     2701 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     2761 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     2821 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2881 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2941 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     3001 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     3061 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     3121 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     3181 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     3241 agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt
     3301 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     3361 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     3421 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     3481 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     3541 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     3601 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     3661 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     3721 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     3781 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     3841 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     3901 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3961 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4021 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     4081 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac
     4141 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     4201 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4261 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4321 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4381 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4441 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4501 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4561 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4621 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4681 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4741 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4801 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4861 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4921 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4981 cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta
     5041 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag
     5101 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     5161 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     5221 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     5281 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     5341 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
     5401 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     5461 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     5521 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     5581 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     5641 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     5701 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     5761 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     5821 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     5881 agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5941 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6001 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6061 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6121 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6181 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6241 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6301 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6361 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6421 gggcgcgtcc cattcgcca
//
