LOCUS       sequence-173055-        5604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DNA-binding (ETS) domain of murine Ets-1, residues 311 to 
            440.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5604)
  AUTHORS   Wang S, Linde MH, Munde M, Carvalho VD, Wilson WD, Poon GM
  TITLE     Mechanistic heterogeneity in site recognition by the structurally 
            homologous DNA-binding domains of the ETS family transcription 
            factors Ets-1 and PU.1.
  JOURNAL   J Biol Chem. 2014 Aug 1;289(31):21605-16. doi: 
            10.1074/jbc.M114.575340. Epub 2014 Jun 21.
  PUBMED    24952944
REFERENCE   2  (bases 1 to 5604)
  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"; doi: 
            "10.1074/jbc.M114.575340"; journalName: "J Biol Chem"; date: 
            "2014-08-1- 1"; volume: "289"; issue: "31"; pages: "21605-16"
COMMENT     Sequence Label: pET28b-Ets1-ETS
FEATURES             Location/Qualifiers
     source          1..5604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1305..1496
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1490..1512)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1598..1737
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1653..1672
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1752..1769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1923..2511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2003..2022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2622..2641)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2633..3448
                     /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             2633..3448
                     /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(2706..2725)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3540..3995)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3677..3698)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3889..3908
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4032..4079)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4075..4093
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4146..4163)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4173..4190)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4191..4208)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             4551..4573
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4588..4612
                     /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(4612..4631)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4613..4631)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4715..4734)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4940..5017
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5018..5604,1..496)
                     /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(5037..5056)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
       61 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
      121 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
      181 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
      241 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
      301 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
      361 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
      421 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
      481 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg
      541 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
      601 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
      661 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
      721 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
      781 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccccac
      841 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
      901 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
      961 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     1021 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     1081 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     1141 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     1201 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     1261 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
     1321 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
     1381 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
     1441 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1501 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1561 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     1621 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     1681 gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1741 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1921 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1981 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2041 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2101 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2161 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2221 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2281 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2341 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2401 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2461 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2521 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2581 gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2641 tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt
     2701 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     2761 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2821 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2881 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2941 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3001 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3061 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3121 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3181 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3241 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3301 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3361 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3421 tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat
     3481 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     3541 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3601 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3661 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3721 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3781 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3841 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3901 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3961 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccaatcc ggatatagtt
     4021 cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt
     4081 tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga
     4141 tctcagtggt ggtggtggtg gtgctcgagt tagtgatggt gatggtgatg ggaacctcgt
     4201 ggcactagct cgtcggcatc tggcttgacg tccagcatgg cgtgcagctc ctcaggggtg
     4261 taccccagca ggctctgcag gtcacacaca aagcggtaca cgtagcgttt ccccgctgtc
     4321 ttgtggatga tgtttttgtc gtaatagtag cgtaggccac ggctcagttt ctcataattc
     4381 atcttaggtt tgtttttcct ctttccccat ctcctggcca cctcatctgg gtcagaaagt
     4441 ttgaattccc agccatctcc tgtccagctg ataaaagact gacaggattt atcagtgagt
     4501 aattccagaa gaaactgcca tagctggatt ggtccactgc ccatggtata tctccttctt
     4561 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
     4621 gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc
     4681 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     4741 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     4801 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     4861 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     4921 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     4981 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     5041 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     5101 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     5161 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     5221 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     5281 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     5341 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     5401 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     5461 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     5521 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     5581 attaagttct gtctcggcgc gtct
//
