LOCUS       sequence-438996-        7525 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 7525)
  AUTHORS   You J, Croyle JL, Nishimura A, Ozato K, Howley PM
  TITLE     Interaction of the bovine papillomavirus E2 protein with Brd4 
            tethers the viral DNA to host mitotic chromosomes.
  JOURNAL   Cell. 2004 Apr 30. 117(3):349-60.
  PUBMED    15109495
REFERENCE   2  (bases 1 to 7525)
  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: "Cell. 2004 
            Apr 30. 117(3):349-60."
COMMENT     Sequence Label: p5071 pcDNA4c hBrd4 (595-1362)
FEATURES             Location/Qualifiers
     source          1..7525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    917..1079
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             1092..1109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1113..1145
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1149..1157,1158..1172)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1614..1631
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3533..3550)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3539..3763
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3809..4237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3896..3915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4106..4127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4246..4266)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4251..4580
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4431..4566
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4493..4512
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4628..4675
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4694..5068
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5198..5319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5235..5254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5289..5308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5368..5384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5368..5384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5381..5403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5392..5408
                     /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(5416..5422,5423..5440,5441..5446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5461..5482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5599..5616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5770..6358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5850..5869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6529..7320,7321..7389))
                     /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     7152..7171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7390..7494)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttagcg cagaggcttg gggcagccga gcggcagcca ggccccggcc
      961 cgggcctcgg ttccagaagg gagaggagcc cgccaaggcg cgcaagagag cgggctgcct
     1021 cgcagtccga gccggagagg gagcgcgagc cgcgccggcc ccggacggcc tccgaaacca
     1081 tggggggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
     1141 tgggtcggga tctgtacgac gatgacgata aggtaccagg atccccacct cccacgtatg
     1201 agtcggagga agaggacaag tgcaagccta tgtcctatga ggagaagcgg cagctcagct
     1261 tggacatcaa caagctcccc ggcgagaagc tgggccgcgt ggtgcacatc atccagtcac
     1321 gggagccctc cctgaagaat tccaaccccg acgagattga aatcgacttt gagaccctga
     1381 agccgtccac actgcgtgag ctggagcgct atgtcacctc ctgtttgcgg aagaaaagga
     1441 aacctcaagc tgagaaagtt gatgtgattg ccggctcctc caagatgaag ggcttctcgt
     1501 cctcagagtc ggagagctcc agtgagtcca gctcctctga cagcgaagac tccgaaacag
     1561 agatggctcc gaagtcaaaa aagaaggggc accccgggag ggagcagaag aagcaccatc
     1621 atcaccacca tcagcagatg cagcaggccc cggctcctgt gccccagcag ccgcccccgc
     1681 ctccccagca gcccccaccg cctccacctc cgcagcagca acagcagccg ccacccccgc
     1741 ctcccccacc ctccatgccg cagcaggcag ccccggcgat gaagtcctcg cccccaccct
     1801 tcattgccac ccaggtgccc gtcctggagc cccagctccc aggcagcgtc tttgacccca
     1861 tcggccactt cacccagccc atcctgcacc tgccgcagcc tgagctgccc cctcacctgc
     1921 cccagccgcc tgagcacagc actccacccc atctcaacca gcacgcagtg gtctctcctc
     1981 cagctttgca caacgcacta ccccagcagc catcacggcc cagcaaccga gccgctgccc
     2041 tgcctcccaa gcccgcccgg cccccagccg tgtcaccagc cttgacccaa acacccctgc
     2101 tcccacagcc ccccatggcc caaccccccc aagtgctgct ggaggatgaa gagccacctg
     2161 ccccacccct cacctccatg cagatgcagc tgtacctgca gcagctgcag aaggtgcagc
     2221 cccctacgcc gctactccct tccgtgaagg tgcagtccca gcccccaccc cccctgccgc
     2281 ccccacccca cccctctgtg cagcagcagc tgcagcagca gccgccacca cccccaccac
     2341 cccagcccca gcctccaccc cagcagcagc atcagccccc tccacggccc gtgcacttgc
     2401 agcccatgca gttttccacc cacatccaac agcccccgcc accccagggc cagcagcccc
     2461 cccatccgcc cccaggccag cagccacccc cgccgcagcc tgccaagcct cagcaagtca
     2521 tccagcacca ccattcaccc cggcaccaca agtcggaccc ctactcaacc ggtcacctcc
     2581 gcgaagcccc ctccccgctt atgatacatt ccccccagat gtcacagttc cagagcctga
     2641 cccaccagtc tccaccccag caaaacgtcc agcctaagaa acaggagctg cgtgctgcct
     2701 ccgtggtcca gccccagccc ctcgtggtgg tgaaggagga gaagatccac tcacccatca
     2761 tccgcagcga gcccttcagc ccctcgctgc ggccggagcc ccccaagcac ccggagagca
     2821 tcaaggcccc cgtccacctg ccccagcggc cggaaatgaa gcctgtggat gtcgggaggc
     2881 ctgtgatccg gcccccagag cagaacgcac cgccaccagg ggcccctgac aaggacaaac
     2941 agaaacagga gccgaagact ccagttgcgc ccaaaaagga cctgaaaatc aagaacatgg
     3001 gctcctgggc cagcctagtg cagaagcatc cgaccacccc ctcctccaca gccaagtcat
     3061 ccagcgacag cttcgagcag ttccgccgcg ccgctcggga gaaagaggag cgtgagaagg
     3121 ccctgaaggc tcaggccgag cacgctgaga aggagaagga gcggctgcgg caggagcgca
     3181 tgaggagccg agaggacgag gatgcgctgg agcaggcccg gcgggcccat gaggaggcac
     3241 gtcggcgcca ggagcagcag cagcagcagc gccaggagca acagcagcag cagcaacagc
     3301 aagcagctgc ggtggctgcc gccgccaccc cacaggccca gagctcccag ccccagtcca
     3361 tgctggacca gcagagggag ttggcccgga agcgggagca ggagcgaaga cgccgggaag
     3421 ccatggcagc taccattgac atgaatttcc agagtgatct attgtcaata tttgaagaaa
     3481 atcttttcgc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     3541 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3601 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3661 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3721 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3781 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3841 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3901 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3961 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4021 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4081 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4141 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4201 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4261 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4321 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4381 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4441 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4501 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4561 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4621 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4681 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     4741 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     4801 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     4861 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     4921 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     4981 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     5041 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     5101 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5161 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5221 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5281 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5341 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5401 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5461 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5521 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5581 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5641 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5701 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5761 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5821 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5881 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5941 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6001 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6061 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6181 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6601 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6721 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6961 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7021 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7141 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7201 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7261 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7321 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7381 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7441 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7501 tccccgaaaa gtgccacctg acgtc
//
