LOCUS       sequence-140248-        7951 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 7951)
  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 7951)
  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: p6348 MSCV-CMV-Flag-HA-Brd4 1047-1362
FEATURES             Location/Qualifiers
     source          1..7951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2026..2044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2082..2104)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2204..2223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2417..2439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2567..2586
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2786..3302
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3366..3707
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3774..4190
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4120..4142
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4158..4174
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        4205..4584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4585..4788
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4739..4759
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4790..4808
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4811..4829
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4839..4863
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4936..4945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4945..4968
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4981..5007
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    5023..5047
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5071..5091
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6059..6083)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6102..6601
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6137..6160)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6534..6553
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6622..7221
                     /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(6622..6641)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7118..7138
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    7807..7823
                     /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(7831..7837,7838..7855,7856..7861))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7876..7897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2041 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2101 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2161 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2221 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2281 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2341 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2401 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat
     2461 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     2521 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     2581 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     2641 agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     2701 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     2761 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     2821 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     2881 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     2941 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     3001 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     3061 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     3121 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     3181 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac
     3241 tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca
     3301 tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg
     3361 gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg
     3421 gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga
     3481 cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg
     3541 actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc
     3601 cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     3661 aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca
     3721 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga
     3781 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
     3841 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
     3901 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
     3961 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
     4021 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
     4081 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
     4141 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat
     4201 ccttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     4261 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     4321 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     4381 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4441 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     4501 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4561 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     4621 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4681 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4741 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     4801 gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag
     4861 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     4921 agcctccggg gatctgccac catggactac aaggatgacg atgacaagct cgatggagga
     4981 tacccatacg atgttccaga ttacgctgct cgaggttatc aaacaagttt gtacaaaaaa
     5041 gcaggctccg cggccgcccc cttcacccct cctaagaaga agaggaaggt tcggcaccac
     5101 aagtcggacc cctactcaac cggtcacctc cgcgaagccc cctccccgct tatgatacat
     5161 tccccccaga tgtcacagtt ccagagcctg acccaccagt ctccacccca gcaaaacgtc
     5221 cagcctaaga aacaggagct gcgtgctgcc tccgtggtcc agccccagcc cctcgtggtg
     5281 gtgaaggagg agaagatcca ctcacccatc atccgcagcg agcccttcag cccctcgctg
     5341 cggccggagc cccccaagca cccggagagc atcaaggccc ccgtccacct gccccagcgg
     5401 ccggaaatga agcctgtgga tgtcgggagg cctgtgatcc ggcccccaga gcagaacgca
     5461 ccgccaccag gggcccctga caaggacaaa cagaaacagg agccgaagac tccagttgcg
     5521 cccaaaaagg acctgaaaat caagaacatg ggctcctggg ccagcctagt gcagaagcat
     5581 ccgaccaccc cctcctccac agccaagtca tccagcgaca gcttcgagca gttccgccgc
     5641 gccgctcggg agaaagagga gcgtgagaag gccctgaagg ctcaggccga gcacgctgag
     5701 aaggagaagg agcggctgcg gcaggagcgc atgaggagcc gagaggacga ggatgcgctg
     5761 gagcaggccc ggcgggccca tgaggaggca cgtcggcgcc aggagcagca gcagcagcag
     5821 cgccaggagc aacagcagca gcagcaacag caagcagctg cggtggctgc cgccgccacc
     5881 ccacaggccc agagctccca gccccagtcc atgctggacc agcagaggga gttggcccgg
     5941 aagcgggagc aggagcgaag acgccgggaa gccatggcag ctaccattga catgaatttc
     6001 cagagtgatc tattgtcaat atttgaagaa aatcttttct gaaagggtgg gcgcgccgac
     6061 ccagctttct tgtacaaagt ggttcgataa cgaattctac cgggtagggg aggcgctttt
     6121 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     6181 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     6241 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     6301 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     6361 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     6421 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     6481 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     6541 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     6601 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     6661 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     6721 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     6781 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     6841 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     6901 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     6961 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     7021 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     7081 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     7141 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     7201 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     7261 gcacgacccc atgcatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     7321 atgaaagacc ccacctgtag gtttggcaag ctagagaacc atcagatgtt tccagggtgc
     7381 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     7441 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     7501 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     7561 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     7621 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     7681 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     7741 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     7801 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7861 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7921 tttccagtcg ggaaacctgt cgtgccagct g
//
