LOCUS       sequence-450974-        5491 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 5491)
  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 5491)
  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: p5081 pcDNA4c hBrd4 (1134-1223)
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..91
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        72..90
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    126..288
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             301..318
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             322..354
                     /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(358..366,367..381)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(708..725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    714..938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      984..1412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1071..1090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1281..1302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1421..1441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1426..1755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1606..1741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1668..1687
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1803..1850
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1869..2243
                     /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    2373..2494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2410..2429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2464..2483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2543..2559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2543..2559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2556..2578)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2567..2583
                     /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(2591..2597,2598..2615,2616..2621))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2636..2657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2774..2791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2945..3533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3025..3044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3704..4495,4496..4564))
                     /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     4327..4346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4565..4669)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4744..4763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4935..5314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5315..5491,1..27)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5469..5489
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
       61 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta
      121 agcttagcgc agaggcttgg ggcagccgag cggcagccag gccccggccc gggcctcggt
      181 tccagaaggg agaggagccc gccaaggcgc gcaagagagc gggctgcctc gcagtccgag
      241 ccggagaggg agcgcgagcc gcgccggccc cggacggcct ccgaaaccat ggggggttct
      301 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
      361 ctgtacgacg atgacgataa ggtaccagga tcccccaagc acccggagag catcaaggcc
      421 cccgtccacc tgccccagcg gccggaaatg aagcctgtgg atgtcgggag gcctgtgatc
      481 cggcccccag agcagaacgc accgccacca ggggcccctg acaaggacaa acagaaacag
      541 gagccgaaga ctccagttgc gcccaaaaag gacctgaaaa tcaagaacat gggctcctgg
      601 gccagcctag tgcagaagca tccgaccacc ccctcctcca cagccaagtc atccagcgac
      661 agcgcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
      721 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
      781 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
      841 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
      901 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
      961 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1021 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1081 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1141 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1201 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1261 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1321 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     1381 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     1441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     1501 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1561 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     1621 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1681 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     1741 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     1801 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     1861 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     1921 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     1981 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     2041 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     2101 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     2161 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     2221 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     2281 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     2341 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     2401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2461 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     2521 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2581 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2641 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2701 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4681 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     4741 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     4801 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     4861 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     4921 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     4981 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     5041 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5101 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5161 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     5221 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     5281 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     5341 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     5401 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     5461 cccattgacg caaatgggcg gtaggcgtgt a
//
