LOCUS       sequence-170057-        5602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses flag tagged Ets1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5602)
  AUTHORS   Seidel JJ, Graves BJ
  TITLE     An ERK2 docking site in the Pointed domain distinguishes a subset of
            ETS transcription factors.
  JOURNAL   Genes Dev. 2002 Jan 1;16(1):127-37.
  PUBMED    11782450
REFERENCE   2  (bases 1 to 5602)
  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: "Genes 
            Dev."; date: "2002-01-1"; volume: "16(1)"; pages: "127-37"
COMMENT     Sequence Label: pCMV-mFlagEts1
FEATURES             Location/Qualifiers
     source          1..5602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        562..580
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             624..647
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2045..2064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2046..2064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2338..2459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2375..2394)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2429..2448
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2466..2921)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2603..2624)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2815..2834
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2948..3052
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3054..3411
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3054..3245
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3077..3097)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3262..3397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3446..4240
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3446..4240
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3500..3519)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4110..4129
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4428..4447)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4472..4519
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4848..5436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5337..5356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      541 tagcgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca
      601 ccgcggtggc ggccgccacc atggattaca aggatgacga cgataaggcc cgggcggatc
      661 ccccgggctg catgaaggcg gccgtcgatc tcaagccgac tctcaccatc atcaagacag
      721 aaaaagtgga tctcgagctt ttcccttccc cggacatgga atgtgcagat gtcccgctgt
      781 taactccgag cagcaaagaa atgatgtccc aagccttgaa agctactttc agtggtttca
      841 caaaagaaca gcagcgactg ggaatcccca aagacccccg gcagtggaca gaaacccacg
      901 tccgggactg ggtgatgtgg gctgtgaatg agttcagcct gaaaggtgtg gacttccaga
      961 agttctgcat gagtggagca gcactgtgtg ccctgggtaa agaatgcttc ctcgagctgg
     1021 ctccagactt tgttggggat atcctgtggg agcatctaga gatcctgcag aaagaggatg
     1081 tgaaaccata tcaggttaat ggagccaacc ctacctaccc agaatcctgt tacacctcgg
     1141 attacttcat cagctacggt atcgagcatg ctcagtgtgt tcctccctca gagttctcag
     1201 agcccagctt catcacagag tcctatcaga cgctgcatcc tatcagctcg gaagaactcc
     1261 tgtccctcaa gtatgagaac gactaccctt ctgtcattct ccaggaccct ctccagacag
     1321 acaccttgca gacagactac tttgccatca agcaagaggt gttaactcca gacaacatgt
     1381 gcctggggag agccagtcgt ggtaaactcg ggggccagga ctcttttgag agcgtagaga
     1441 gctacgatag ttgtgaccgc ctcacccagt cctggagcag ccagtcatcc ttcaacagcc
     1501 tgcagcgggt cccctcctat gacagcttcg actacgagga ttatcccgct gccctgccca
     1561 accacaagcc caagggcacc ttcaaggact atgtgcgtga ccgtgctgac ctcaacaagg
     1621 acaagcctgt cattcctgct gctgccctgg ctggctacac aggaagtggg ccgatccagc
     1681 tgtggcagtt tcttctggaa ttactcactg ataagtcttg tcagtccttt atcagctgga
     1741 caggagatgg ctgggaattc aagctttctg acccagatga ggtggccagg agatggggaa
     1801 agaggaaaaa caaacctaag atgaattatg agaaactgag ccgtggcctt cgctactatt
     1861 atgacaaaaa tatcatccac aagacggcgg gcaagcgcta cgtataccgc tttgtgtgcg
     1921 acctgcagag cctgctggga tacacccctg aagagctgca cgccatgctg gatgtaaagc
     1981 cggatgctga ctaggggccc ggtaccttaa ttaattaagg taccaggtaa gtgtacccaa
     2041 ttcgccctat agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag
     2101 cctgaatggc gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta
     2161 attgtttgtg tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac
     2221 agtctgttca tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2281 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2341 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2401 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2461 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2521 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2581 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2641 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2701 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2761 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2821 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2881 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     2941 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3001 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga atcctgaggc
     3061 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3241 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3301 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3361 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3421 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3481 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3541 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3601 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3661 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3721 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3781 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3841 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3901 tgtcgatcag gatgatctgg acgaagaaca tcaggggctc gcgccagccg aactgttcgc
     3961 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4021 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4081 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagaact
     4141 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4201 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4261 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4321 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4381 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4441 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4501 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4561 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4621 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4681 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4981 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5521 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     5581 attagttcat agcccatata tg
//
