LOCUS       sequence-258766-        8137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8137)
  AUTHORS   Rahman S, Sowa ME, Ottinger M, Smith JA, Shi Y, Harper JW, Howley PM
  TITLE     The Brd4 Extraterminal Domain Confers Transcription Activation 
            Independent of pTEFb by Recruiting Multiple Proteins, Including 
            NSD3.
  JOURNAL   Mol Cell Biol. 2011 Jul . 31(13):2641-52.
  PUBMED    21555454
REFERENCE   2  (bases 1 to 8137)
  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: "Mol Cell 
            Biol. 2011 Jul . 31(13):2641-52."
COMMENT     Sequence Label: p6352 MSCV-CMV-CMV-Flag-HA-JMJD6
FEATURES             Location/Qualifiers
     source          1..8137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1420..1799
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1800..2003
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1954..1974
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2005..2023
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2026..2044
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2054..2078
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2151..2160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2160..2183
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2196..2222
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2238..2262
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2825..2842)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     protein_bind    complement(3472..3496)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3515..4002
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3550..3573)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3935..3954
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4023..4622
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4023..4042)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4519..4539
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    5208..5224
                     /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)."
     promoter        complement(join(5232..5238,5239..5256,5257..5262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5277..5298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5415..5432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5586..6174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5666..5685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6345..7136,7137..7205))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6968..6987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7206..7310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7378..7396
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7434..7456)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7556..7575
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7769..7791
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7919..7938
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatccttg acattgatta ttgactagtt
     1441 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1501 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1561 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1621 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1681 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1741 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1801 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1861 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1921 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1981 gggaggtcta tataagcaga gctctcccta tcagtgatag agatctccct atcagtgata
     2041 gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     2101 gttttgacct ccatagaaga caccgggacc gatccagcct ccggggatct gccaccatgg
     2161 actacaagga tgacgatgac aagctcgatg gaggataccc atacgatgtt ccagattacg
     2221 ctgctcgagg ttatcaaaca agtttgtaca aaaaagttgg catgaaccac aagagcaaga
     2281 agcgcatccg cgaggccaag cggagtgcgc ggccggagct caaggactcg ctggattgga
     2341 cccggcacaa ctactacgag agcttctcgc tgagcccggc ggccgtggcg gataacgtgg
     2401 aaagggcaga tgctttacag ctgtctgtgg aagaatttgt ggagcggtat gaaagacctt
     2461 acaagcccgt ggttttgttg aatgcgcaag agggctggtc tgcgcaggag aaatggactc
     2521 tggagcgcct aaaaaggaaa tatcggaacc agaagttcaa gtgtggtgag gataacgatg
     2581 gctactcagt gaagatgaag atgaaatact acatcgagta catggagagc actcgagatg
     2641 atagtcccct ttacatcttt gacagcagct atggtgaaca ccctaaaaga aggaaacttt
     2701 tggaagacta caaggtgcca aagtttttca ctgatgacct tttccagtat gctggggaga
     2761 agcgcaggcc cccttacagg tggtttgtga tggggccacc acgctccgga actgggattc
     2821 acatcgaccc tctgggaacc agtgcctgga atgccttagt tcagggccac aagcgctggt
     2881 gcctgtttcc taccagcact cccagggaac tcatcaaagt gacccgagac gaaggaggga
     2941 accagcaaga cgaagctatt acctggttta atgttattta tccccggaca cagcttccaa
     3001 cctggccacc tgaattcaaa cccctggaaa tcttacaaaa accaggagag actgtctttg
     3061 taccaggagg ctggtggcat gttgtcctca atctcgacac tactatcgcc atcacccaaa
     3121 attttgccag cagcaccaac ttccctgtgg tatggcacaa gacggtaaga gggagaccaa
     3181 agttatcaag gaaatggtat aggattttga agcaagagca ccccgagttg gcagtcctcg
     3241 cagactcggt tgaccttcag gagtccacag ggatagcttc cgacagctcc agcgactctt
     3301 ccagctcctc cagctccagt tcgtcagact ccgactcaga gtgcgagtct ggatccgagg
     3361 gcgatgggac agtgcaccgc aggaagaaga ggaggacgtg cagcatggtg ggaaacgggg
     3421 acaccacctc ccaggacgac tgtgtcagca aagagcgcag ctcctccagg tgcccaactt
     3481 tcttgtacaa agtggttcga taacgaattc taccgggtag gggaggcgct tttcccaagg
     3541 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     3601 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     3661 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     3721 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     3781 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     3841 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     3901 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     3961 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     4021 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     4081 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     4141 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4201 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4261 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4321 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4381 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4441 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4501 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4561 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4621 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
     4681 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4741 ggtttggcaa gctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     4801 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     4861 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     4921 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     4981 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     5041 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata
     5101 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa
     5161 atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc
     5221 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5281 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5341 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6481 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7321 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7381 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     7441 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     7501 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     7561 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     7621 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     7681 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     7741 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa
     7801 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     7861 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     7921 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     7981 tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg
     8041 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     8101 aataaaagat tttatttagt ctccagaaaa agggggg
//
