LOCUS       sequence-175402-        3740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DNA-binding (ETS) domain of murine PU.1, residues 167-272,
            in M15 cells already pre-transformed with 2nd plasmid, pREP-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3740)
  AUTHORS   Poon GM
  TITLE     Sequence discrimination by DNA-binding domain of ETS family 
            transcription factor PU.1 is linked to specific hydration of 
            protein-DNA interface.
  JOURNAL   J Biol Chem. 2012 May 25;287(22):18297-307. doi: 
            10.1074/jbc.M112.342345. Epub 2012 Apr 2.
  PUBMED    22474303
REFERENCE   2  (bases 1 to 3740)
  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.M112.342345"; journalName: "J Biol Chem"; date: 
            "2012-05-25- 25"; volume: "287"; issue: "22"; pages: "18297-307"
COMMENT     Sequence Label: pQE60-mPU.1-ETS
FEATURES             Location/Qualifiers
     source          1..3740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /gene="lacI"
                     /label=lacI promoter
     CDS             79..1161
                     /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(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(1539..2333)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1650..1669)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2260..2279
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      2820..3365
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3266..3285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3482..3499
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg aattgtcgac
     1201 caaagcggcc atcgtgcctc cccactcctg cagttcgggg gcatggatgc gcggatagcc
     1261 gctgctggtt tcctggatgc cgacggattt gcactgccgg tagaactccg cgaggtcgtc
     1321 cagcctcagg cagcagctga accaactcgc gaggggatcg agcccggggt gggcgaagaa
     1381 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     1441 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     1501 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     1561 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     1621 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     1681 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     1741 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     1801 atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     1861 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     1921 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     1981 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     2041 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     2101 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca
     2161 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     2221 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     2281 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaag catgcgaaac
     2341 gaccgtcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc
     2401 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
     2461 ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc
     2521 ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag
     2581 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgttccgc
     2641 ttcctttagc agcccttgcg ccctgagtgc ttgcggcagc gtgaagctta tcgatgataa
     2701 gctgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt
     2761 gaagagataa attgcactga aatctagaaa tattttatct gattaataag atgatcttct
     2821 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     2881 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     2941 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     3001 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     3061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     3121 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     3181 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     3241 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3301 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     3361 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     3421 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     3481 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     3541 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     3601 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
     3661 gcaccacccc gtcagtagct gaacaggagg gacagggtcg acaattcgcg cgcgaaggcg
     3721 aagcggcatg catttacgtt
//
