LOCUS       sequence-140215-        7840 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 7840)
  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 7840)
  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: p6347 MSCV-CMV-Flag-HA-Brd4-444-722
FEATURES             Location/Qualifiers
     source          1..7840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     CDS             complement(783..1382)
                     /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(866..886)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1363..1382
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1403..1902)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1451..1470)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1844..1867
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    1921..1945
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2802..2822)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(2846..2870)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2886..2912)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2925..2948)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2948..2957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3030..3054)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3064..3082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3085..3103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3105..3308)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3134..3154)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3309..3688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3703..4119)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3719..3735)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3751..3773)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4186..4527
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4591..5107
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5307..5326)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5454..5476)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5670..5689)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5789..5811
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5849..5867)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5935..6039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6040..6108,6109..6900)
                     /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     complement(6258..6277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7071..7659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7560..7579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7813..7830
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
      241 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
      301 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
      361 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      421 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
      481 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
      541 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      601 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagcttgcca
      661 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
      721 tttatcgatg catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc gtggggcggg
      781 cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
      841 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
      901 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
      961 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     1021 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     1081 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     1141 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     1201 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     1261 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     1321 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     1381 atggtaagct tgggctgcag gtcgaaaggc ccggagatga ggaagaggag aacagcgcgg
     1441 cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt cgggcgcccg
     1501 ccccgcccct gagcccgccc ctgagcccgc ccccggaccc accccttccc agcctctgag
     1561 cccagaaagc gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct acccgcttcc
     1621 attgctcagc ggtgctgtcc atctgcacga gactagtgag acgtgctact tccatttgtc
     1681 acgtcctgca cgacgcgagc tgcggggcgg gggggaactt cctgactagg ggaggagtag
     1741 aaggtggcgc gaaggggcca ccaaagaacg gagccggttg gcgcctaccg gtggatgtgg
     1801 aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt gcccagcggg gctgctaaag
     1861 cgcatgctcc agactgcctt gggaaaagcg cctcccctac ccggtagaat tcgttatcga
     1921 accactttgt acaagaaagc tgggtcggcg cgcccaccct tttaggcagg acctgtttcg
     1981 gagtcttcgc tgtcagagga gctggactca ctggagctct ccgactctga ggacgagaag
     2041 cccttcatct tggaggagcc ggcaatcaca tcaactttct cagcttgagg tttccttttc
     2101 ttccgcaaac aggaggtgac atagcgctcc agctcacgca gtgtggacgg cttcagggtc
     2161 tcaaagtcga tttcaatctc gtcggggttg gaattcttca gggagggctc ccgtgactgg
     2221 atgatgtgca ccacgcggcc cagcttctcg ccggggagct tgttgatgtc caagctgagc
     2281 tgccgcttct cctcatagga cataggcttg cacttgtcct cttcctccga ctcatacgtg
     2341 ggagggggct tgctcttcat gggcgctggc tccttcttgc tcacattgct gttgctgcta
     2401 ttatttttct tcgtcttttt aggaggaggt tccttggctt tgcttttttt attctcttcc
     2461 acttcctctt tccttttgtg cttttctttt ttcttttcct tcttgtcttt ctcctttttc
     2521 tttggtttgt tctgctgggg ctgagagagg gctgcaagct gctcgtgcac ggctttgagc
     2581 tgctcctgga gctcagccag ccgctgggct cgctcctcct cagagtcatc agtcgaactg
     2641 tcactgtccg aggagctatc gctgctgctg tcgctggatg agggcggggc cacaaccttg
     2701 gtgggagggg gcactgccgg ggaggacacg gccaccactg gctcctcagg ctcgtccggc
     2761 atcttggcaa agcgcatttc gaacacatcc tggagcttgc gcaccttcct cttcttctta
     2821 ggaggggtga agggggcggc cgcggagcct gcttttttgt acaaacttgt ttgataacct
     2881 cgagcagcgt aatctggaac atcgtatggg tatcctccat cgagcttgtc atcgtcatcc
     2941 ttgtagtcca tggtggcaga tccccggagg ctggatcggt cccggtgtct tctatggagg
     3001 tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg agctcgtcga
     3061 cgatctctat cactgatagg gagatctcta tcactgatag ggagagctct gcttatatag
     3121 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     3181 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     3241 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     3301 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     3361 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     3421 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     3481 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     3541 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     3601 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     3661 tattaataac tagtcaataa tcaatgtcaa ggatctaatt ccggcgccta gagaaggagt
     3721 gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa gggggagaga
     3781 cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct tgacccaggg
     3841 aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt ctgggtgtcc
     3901 atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc ggttaaaggt
     3961 gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga aggtaaccca
     4021 acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat ctttccagtg
     4081 acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag acaaatacag
     4141 aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca gacaagacgc
     4201 gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact gttttaggtt
     4261 ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg actcccttcc
     4321 tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag ggttgcggcc
     4381 gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct agttagctaa
     4441 ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa agacagagac
     4501 agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg gtccctgggc
     4561 aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg ggcagtcaat
     4621 caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca aacagcaaga
     4681 ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact ggcgcgccga
     4741 gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga
     4801 gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac
     4861 cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat ctggggacca
     4921 tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca
     4981 tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc agttatgtat
     5041 tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc
     5101 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct atggctcgta
     5161 ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga ccactgatat
     5221 cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg ccggcatcac
     5281 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     5341 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     5401 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc cgtcgtttta
     5461 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5521 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5581 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5641 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5701 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5761 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5821 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5881 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5941 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6001 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6061 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6121 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6181 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6241 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6301 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6361 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6421 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6481 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6541 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6601 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6661 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6721 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6781 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6841 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6901 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6961 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7021 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7141 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7201 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7261 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7321 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7381 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7441 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7501 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7561 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7621 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7681 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7741 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7801 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
