LOCUS       sequence-140288-        8309 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 8309)
  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 8309)
  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: p6346 MSCV-CMV-Flag-HA-Brd4-1-444
FEATURES             Location/Qualifiers
     source          1..8309
                     /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"
     protein_bind    complement(3315..3339)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3355..3381)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3394..3417)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3417..3426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3499..3523)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3533..3551
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3554..3572
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3574..3777)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3603..3623)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3778..4157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4172..4588)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4188..4204)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4220..4242)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4655..4996
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             5060..5576
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5776..5795)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5923..5945)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6139..6158)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6258..6280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6318..6336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6404..6508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6509..6577,6578..7369)
                     /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(6727..6746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7540..8128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8029..8048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8282..8299
                     /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 tagcgggcca tggccaccac
     1981 ctcatggtca ggagggttgt acttatagca gttggagaac atcaatcgga cgtcagcacc
     2041 aaactcctga gcatcacggt actcacgggc ctccagttta gacttgattg tgctcatgtc
     2101 catggggtgc ttgatgatgt cacagtagtc gtgtaggccc agtgcctcca cgtccacagg
     2161 cttgtagaag ggccaggcgt aggcggcgtg cttcttggca aacatctcct tgaggatgcc
     2221 gctgcagcac ttgagctgct ccgagacctt gctgctcttc tctggtgctg ggtgctgctg
     2281 agagtcgggc acgtccttct ttggaggttt cacaggccgg ctgctctccc gccgctggcc
     2341 cagcttggtg gtcttgggct ccgggggcag cgagggtggc tcgtgaatgg ggtcaatggt
     2401 ggtgggggtg gtggtgtctg ctttcctctt cactcccttc tttgtcttca caggctgtgg
     2461 ggtggccgcg atgatgggtg ggtggctctg tacgggctgg ggagctggag cgggtggggg
     2521 ttgtggctgg gggggcactg gcgggggcgt ctgcagtggc tggggaggca ccactgtcat
     2581 gacaggggtc tggacgatga ggtccggggt gacggcaggg aaggggtgag gcgtggcctg
     2641 cacaggagga ggattcggct gaggggtctg ggtctgcgga ggagtcgatg cttgagttgt
     2701 gtttggtacc gtggaaacgc caggttttgc tgtccctgtt tctttcctcc cacgtcctct
     2761 tccttttgcc tggactatca tgatctcggt ttcttctgtg ggtagctcat ttattttttg
     2821 caagaagagc ttttccagag cttctgccat taagactatg tcatctccag gcttgttgta
     2881 gatgtaacaa tttgtaaaca tagtgttgaa gtcctggata cattcctgag cattccagta
     2941 atagttgttt tccaagcgct tctttattgt tcccatatcc ataggcgttt taatgatctt
     3001 atagtaatca gggaggttca gcttgacggc atccacaggc tgctggaaag gccatgcaaa
     3061 ctggtgtttc catagtgtct tgagcaccac tctgagcagg tattgcagtt ggttggtctg
     3121 cctcttgggc ttgttagggt tggaggtctc tgggggcggg gggttggtgc tggctgcgtt
     3181 ggctggctgg ggttgggcct gggcctgtgt tgtagacatt tgggaagttt ctagtccatc
     3241 ccccattact ggcagatttc tcaatctcgt cccagggccg ctctccgcag acatggtgaa
     3301 gggggcggcc gcggagcctg cttttttgta caaacttgtt tgataacctc gagcagcgta
     3361 atctggaaca tcgtatgggt atcctccatc gagcttgtca tcgtcatcct tgtagtccat
     3421 ggtggcagat ccccggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg
     3481 tggatggcgt ctccaggcga tctgacggtt cactaaacga gctcgtcgac gatctctatc
     3541 actgataggg agatctctat cactgatagg gagagctctg cttatataga cctcccaccg
     3601 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     3661 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     3721 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
     3781 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     3841 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     3901 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     3961 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     4021 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     4081 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     4141 agtcaataat caatgtcaag gatctaattc cggcgcctag agaaggagtg agggctggat
     4201 aaagggagga tcgaggcggg gtcgaacgag gaggttcaag ggggagagac ggggcggatg
     4261 gaggaagagg aggcggaggc ttagggtgta caaagggctt gacccaggga ggggggtcaa
     4321 aagccaaggc ttcccaggtc acgatgtagg ggacctggtc tgggtgtcca tgcgggccag
     4381 gtgaaaagac cttgatctta acctgggtga tgaggtctcg gttaaaggtg ccgtctcgcg
     4441 gccatccgac gttaaaggtt ggccattctg cagagcagaa ggtaacccaa cgtctcttct
     4501 tgacatctac cgactggttg tgagcgatcc gctcgacatc tttccagtga cctaaggtca
     4561 aacttaaggg agtggtaaca gtctggccct aattttcaga caaatacaga aacacagtca
     4621 gacagagaca acacagaacg atgctgcagc gctgcagcag acaagacgcg cggcttcggt
     4681 tccaaaccga aagcaaaaat tcagacggag gcgggaactg ttttaggttc tcgtctccta
     4741 ccagaaccac atatcctgac ggggtcggat tccacatcga ctcccttcct caggtcgggc
     4801 cacaaaaacg gcccccaaag tccctgggac gtctcccagg gttgcggccg ggtgttcaga
     4861 actcgtcagt tccaccacgg gtccgccaga tacagagcta gttagctaac tagtacagac
     4921 gcaggcgcaa acattagatg ccggcacaaa cacgcacaaa gacagagaca gacacgaaac
     4981 gaccgctggc cagcttacct cccggcgggg gggtcggtgg tccctgggca ggggtctcca
     5041 aatctcggtg gaacctccaa atgaaagacc cccgaggtgg gcagtcaatc aatctgagga
     5101 gaccctccca aggatcagcg agtccacgat tcggatgcaa acagcaagag gctttattgg
     5161 gaatacgggt acccgggcga cgcagtctat cggaggactg gcgcgccgag tgaggggttg
     5221 tgggctcttt tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg
     5281 attggttagt tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat
     5341 ctgatggttc tctagaaact gctgagggcg ggaccgcatc tggggaccat ctgttcttgg
     5401 ccctgagccg gggcaggaac tgcttaccac agatatcctg tttggcccat attctgctgt
     5461 ctctctgttc ctaaccttga tctgaacttc tctattctca gttatgtatt ttccatgcct
     5521 tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc
     5581 cctttttctg gagactaaat aaaatctttt attttatcta tggctcgtac tctataggct
     5641 tcagctggtg atattgttga gtcaaaacta gagcctggac cactgatatc ctgtctttaa
     5701 caaattggac taatcgcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     5761 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     5821 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     5881 tgttgggcgc catctccttg catgcaccat tccttgcgcc gtcgttttac aacgtcgtga
     5941 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6001 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6061 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6121 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6181 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6241 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6301 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6361 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6481 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6541 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6601 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6661 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6721 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6781 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6841 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6901 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6961 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7021 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7081 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7141 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7201 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7261 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7321 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7381 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7441 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7501 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7561 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7621 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7681 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7741 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7801 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7861 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7921 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7981 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8041 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8101 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8161 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     8221 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8281 cagcgagtca gtgagcgagg aagcggaag
//
