LOCUS       sequence-147547-        8729 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 8729)
  AUTHORS   Bisgrove DA, Mahmoudi T, Henklein P, Verdin E
  TITLE     Conserved P-TEFb-interacting domain of BRD4 inhibits HIV 
            transcription.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Aug 21. 104(34):13690-5.
  PUBMED    17690245
REFERENCE   2  (bases 1 to 8729)
  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: "Proc Natl 
            Acad Sci U S A. 2007 Aug 21. 104(34):13690-5."
COMMENT     Sequence Label: pFlag-CMV2-Brd4 (1-1362)
FEATURES             Location/Qualifiers
     source          1..8729
                     /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
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2580..2597
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2581..2597
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2758..3137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3138..3341
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3292..3312
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3375..3398
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5613..5630
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7510..8132
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(7709..7728)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(8156..8176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8161..8490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8341..8476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8528..8547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8529..8547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8561..8577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8561..8577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8574..8596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8585..8601
                     /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(8609..8615,8616..8633,8634..8639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8654..8675
                     /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 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2521 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2581 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
     2641 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     2701 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     2761 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2821 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2881 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2941 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3001 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3061 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     3121 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     3181 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3241 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     3301 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     3361 attgatctac catggactac aaagacgatg acgacaagct tatgtctgcg gagagcggcc
     3421 ctgggacgag attgagaaat ctgccagtaa tgggggatgg actagaaact tcccaaatgt
     3481 ctacaacaca ggcccaggcc caaccccagc cagccaacgc agccagcacc aaccccccgc
     3541 ccccagagac ctccaaccct aacaagccca agaggcagac caaccaactg caatacctgc
     3601 tcagagtggt gctcaagaca ctatggaaac accagtttgc atggcctttc cagcagcctg
     3661 tggatgccgt caagctgaac ctccctgatt actataagat cattaaaacg cctatggata
     3721 tgggaacaat aaagaagcgc ttggaaaaca actattactg gaatgctcag gaatgtatcc
     3781 aggacttcaa cactatgttt acaaattgtt acatctacaa caagcctgga gatgacatag
     3841 tcttaatggc agaagctctg gaaaagctct tcttgcaaaa aataaatgag ctacccacag
     3901 aagaaaccga gatcatgata gtccaggcaa aaggaagagg acgtgggagg aaagaaacag
     3961 ggacagcaaa acctggcgtt tccacggtac caaacacaac tcaagcatcg actcctccgc
     4021 agacccagac ccctcagccg aatcctcctc ctgtgcaggc cacgcctcac cccttccctg
     4081 ccgtcacccc ggacctcatc gtccagaccc ctgtcatgac agtggtgcct ccccagccac
     4141 tgcagacgcc cccgccagtg cccccccagc cacaaccccc acccgctcca gctccccagc
     4201 ccgtacagag ccacccaccc atcatcgcgg ccaccccaca gcctgtgaag acaaagaagg
     4261 gagtgaagag gaaagcagac accaccaccc ccaccaccat tgaccccatt cacgagccac
     4321 cctcgctgcc cccggagccc aagaccacca agctgggcca gcggcgggag agcagccggc
     4381 ccgtgaaacc tccaaagaag gacgtgcccg actctcagca gcacccagca ccagagaaga
     4441 gcagcaaggt ctcggagcag ctcaagtgct gcagcggcat cctcaaggag atgtttgcca
     4501 agaagcacgc cgcctacgcc tggcccttct acaagcctgt ggacgtggag gcactgggcc
     4561 tacacgacta ctgtgacatc atcaagcacc ccatggacat gagcacaatc aagtctaaac
     4621 tggaggcccg tgagtaccgt gatgctcagg agtttggtgc tgacgtccga ttgatgttct
     4681 ccaactgcta taagtacaac cctcctgacc atgaggtggt ggccatggcc cgcaagctcc
     4741 aggatgtgtt cgaaatgcgc tttgccaaga tgccggacga gcctgaggag ccagtggtgg
     4801 ccgtgtcctc cccggcagtg ccccctccca ccaaggttgt ggccccgccc tcatccagcg
     4861 acagcagcag cgatagctcc tcggacagtg acagttcgac tgatgactct gaggaggagc
     4921 gagcccagcg gctggctgag ctccaggagc agctcaaagc cgtgcacgag cagcttgcag
     4981 ccctctctca gccccagcag aacaaaccaa agaaaaagga gaaagacaag aaggaaaaga
     5041 aaaaagaaaa gcacaaaagg aaagaggaag tggaagagaa taaaaaaagc aaagccaagg
     5101 aacctcctcc taaaaagacg aagaaaaata atagcagcaa cagcaatgtg agcaagaagg
     5161 agccagcgcc catgaagagc aagccccctc ccacgtatga gtcggaggaa gaggacaagt
     5221 gcaagcctat gtcctatgag gagaagcggc agctcagctt ggacatcaac aagctccccg
     5281 gcgagaagct gggccgcgtg gtgcacatca tccagtcacg ggagccctcc ctgaagaatt
     5341 ccaaccccga cgagattgaa atcgactttg agaccctgaa gccgtccaca ctgcgtgagc
     5401 tggagcgcta tgtcacctcc tgtttgcgga agaaaaggaa acctcaagct gagaaagttg
     5461 atgtgattgc cggctcctcc aagatgaagg gcttctcgtc ctcagagtcg gagagctcca
     5521 gtgagtccag ctcctctgac agcgaagact ccgaaacaga gatggctccg aagtcaaaaa
     5581 agaaggggca ccccgggagg gagcagaaga agcaccatca tcaccaccat cagcagatgc
     5641 agcaggcccc ggctcctgtg ccccagcagc cgcccccgcc tccccagcag cccccaccgc
     5701 ctccacctcc gcagcagcaa cagcagccgc cacccccgcc tcccccaccc tccatgccgc
     5761 agcaggcagc cccggcgatg aagtcctcgc ccccaccctt cattgccacc caggtgcccg
     5821 tcctggagcc ccagctccca ggcagcgtct ttgaccccat cggccacttc acccagccca
     5881 tcctgcacct gccgcagcct gagctgcccc ctcacctgcc ccagccgcct gagcacagca
     5941 ctccacccca tctcaaccag cacgcagtgg tctctcctcc agctttgcac aacgcactac
     6001 cccagcagcc atcacggccc agcaaccgag ccgctgccct gcctcccaag cccgcccggc
     6061 ccccagccgt gtcaccagcc ttgacccaaa cacccctgct cccacagccc cccatggccc
     6121 aaccccccca agtgctgctg gaggatgaag agccacctgc cccacccctc acctccatgc
     6181 agatgcagct gtacctgcag cagctgcaga aggtgcagcc ccctacgccg ctactccctt
     6241 ccgtgaaggt gcagtcccag cccccacccc ccctgccgcc cccaccccac ccctctgtgc
     6301 agcagcagct gcagcagcag ccgccaccac ccccaccacc ccagccccag cctccacccc
     6361 agcagcagca tcagccccct ccacggcccg tgcacttgca gcccatgcag ttttccaccc
     6421 acatccaaca gcccccgcca ccccagggcc agcagccccc ccatccgccc ccaggccagc
     6481 agccaccccc gccgcagcct gccaagcctc agcaagtcat ccagcaccac cattcacccc
     6541 ggcaccacaa gtcggacccc tactcaaccg gtcacctccg cgaagccccc tccccgctta
     6601 tgatacattc cccccagatg tcacagttcc agagcctgac ccaccagtct ccaccccagc
     6661 aaaacgtcca gcctaagaaa caggagctgc gtgctgcctc cgtggtccag ccccagcccc
     6721 ttgtggtggt gaaggaggag aagatccact cacccatcat ccgcagcgag cccttcagcc
     6781 cctcgctgcg gccggagccc cccaagcacc cggagagcat caaggccccc gtccacctgc
     6841 cccagcggcc ggaaatgaag cctgtggatg tcgggaggcc tgtgatccgg cccccagagc
     6901 agaacgcacc gccaccaggg gcccctgaca aggacaaaca gaaacaggag ccgaagactc
     6961 cagttgcgcc caaaaaggac ctgaaaatca agaacatggg ctcctgggcc agcctagtgc
     7021 agaagcatcc gaccaccccc tcctccacag ccaagtcatc cagcgacagc ttcgagcagt
     7081 tccgccgcgc cgctcgggag aaagaggagc gtgagaaggc cctgaaggct caggccgagc
     7141 acgctgagaa ggagaaggag cggctgcggc aggagcgcat gaggagccga gaggacgagg
     7201 atgcgctgga gcaggcccgg cgggcccatg aggaggcacg tcggcgccag gagcagcagc
     7261 agcagcagcg ccaggagcaa cagcagcagc agcaacagca agcagctgcg gtggctgccg
     7321 ccgccacccc acaggcccag agctcccagc cccagtccat gctggaccag cagagggagt
     7381 tggcccggaa gcgggagcag gagcgaagac gccgggaagc catggcagct accattgaca
     7441 tgaatttcca gagtgatcta ttgtcaatat ttgaagaaaa tcttttctga gcgcacctag
     7501 gtggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     7561 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     7621 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     7681 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     7741 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     7801 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     7861 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     7921 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     7981 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     8041 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     8101 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat
     8161 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     8221 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     8281 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     8341 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     8401 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     8461 ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa
     8521 gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt
     8581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     8641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     8701 tccagtcggg aaacctgtcg tgccagctg
//
