LOCUS       sequence-272908-        7305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BcRF1-ORF24 chimera with a C-terminal 2xStrep tag in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  AUTHORS   Castaneda AF, Didychuk AL, Louder RK, McCollum CO, Davis ZH, Nogales
            E, Glaunsinger BA
  TITLE     The gammaherpesviral TATA-box-binding protein directly interacts 
            with the CTD of host RNA Pol II to direct late gene transcription.
  JOURNAL   PLoS Pathog. 2020 Sep 4;16(9):e1008843. doi: 
            10.1371/journal.ppat.1008843. eCollection 2020 Sep.
  PUBMED    32886723
REFERENCE   2  (bases 1 to 7305)
  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: "PLoS 
            Pathog."; date: "2020-09-4"; volume: "16(9):e1008843. doi"; pages: "
            10.1371/journal.ppat.1008843. eCollection 2020 Sep"
COMMENT     Sequence Label: pcDNA4/TO-BcRF1-ORF24-2xStrep
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2970..2993
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3030..3053
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3079..3096)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3085..3309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3355..3783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3442..3461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3652..3673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3792..3812)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3797..4126
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3977..4112
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4039..4058
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4174..4221
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4240..4614
                     /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    4744..4865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4781..4800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4835..4854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4914..4930)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4914..4930)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4927..4949)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4938..4954
                     /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(4962..4968,4969..4986,4987..4992))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5007..5028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5145..5162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5316..5904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5396..5415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6075..6866,6867..6935))
                     /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     6698..6717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6936..7040)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7115..7134)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccatg acacaaggta
      781 agagggagat ggggggaggt ctcgagggct tctcttcaca gctgggtctc ttttacgccc
      841 tggcctgcaa ccgcagccca cctgcacttc ccgaggatgc tacccttcta atcaaatggt
      901 tggacacggc cctgggcagg gaggccacct tttacgcgtg tcgggctatg cgtcgtcttc
      961 tactcggcgt tatccgaata aatgactgcc aggagctgcc acccggttta ataattctga
     1021 gtccgggcac cgtccctggc ccccttggag tccagagtct ggagcataca gactgcgaaa
     1081 tatggtcctc tgcccaccct gaccacgctg cccacctccc ggtgcccagg gtcatcacat
     1141 acaccgactg cccgggttcc ataagcacga gctcaatgtt tcgccttatc atccgctact
     1201 tgtctcatca ccaatttgag cgctgcttcg agcagttctg ccgcgtggtc ccgcgtcgct
     1261 tcctagggac ctgtaagcaa aactctgcaa agatgctggc tagcctgaag catctctcgt
     1321 tttcaataat gccacccgca tccccagacg cacagctaca ttttttaaag tttaacatcg
     1381 gcagcttcat ggccacgtgg gggtggcacg gagagctggt ctcgctgcgc cgtgccatcg
     1441 ctcacaacgt agagcgactg cccaccgtgc tgaagaacct gtcgaaacag agtaagcacc
     1501 aggacgtcaa ggttaacgga cgggatctgg tgggctttca gctggctcta aaccagctcg
     1561 tgtcccgtct gcacgtaaaa atccaacgca aggaccccgg accaaagcca tacagggtgg
     1621 tcgtcagtac cccagattgt acctactatc tagtgtatcc gggcacaccg gccatctaca
     1681 gactcgtcat gtgtatggca gtggcagact gcatcggcca ctcgtgcagc ggactgcacc
     1741 cctgcgcaaa ctttttaggc acccacgaga caccgcgtct cctggcggcg acgctttcaa
     1801 gaatccggta cgcgccgaaa gaccggcgag cagccatgaa gggaaatttg caggcgtgct
     1861 tccaacgata cgcggccacg gacgcgcgga ctctgggcag ctccacagtg tcagacatgc
     1921 tggaacccac aaaacacgtc agtttggaaa acttcaagat caccatattc aacaccaaca
     1981 tggtgattaa cactaagata agctgccacg ttcctaacac cctgcaaaag actattttaa
     2041 acatccccag attgaccaac aattttgtta tacgaaagta ctccgtaaag gaaccttctt
     2101 ttaccataag cgtgtttttt tccgacaaca tgtgtcaagg caccgcaata aacatcaaca
     2161 tcagtgggga catgctgcac tttctcttcg caatgggtac gctgaaatgc tttctgccaa
     2221 tcaggcacat atttcctgta tcgatagcaa attggaactc cacgttggac ctgcacggac
     2281 tggaaaacca gtacatggtg agaatggggc gaaaaaacgt attttggacc acaaactttc
     2341 catctgtggt ctccagcaag gatgggctaa acgtgtcctg gtttaaggcc gcgacagcca
     2401 cgatttctaa agtgtacggg cagcctcttg tggaacagat tcgccacgag ctggcgccca
     2461 ttctcacgga ccagcacgcg cgcatcgacg gaaacaaaaa tagaatattc tccctacttg
     2521 agcacagaaa ccgttcccaa atacagacgc tacacaaaag gttcctggag tgtctggtgg
     2581 aatgctgttc gtttctcagg cttgacgtgg cttgcattag gcgagccgcc gcccggggcc
     2641 tgtttgactt ctcaaagaag ataatcagtc acactaaaag caaacacgag tgcgcagtac
     2701 tgggatataa aaagtgtaac ctaatcccga aaatctatgc ccgaaacaag aagaccaggc
     2761 tagacgagtt gggccgcaat gcaaacttca tttcgttcgt cgccaccacg ggtcatcggt
     2821 tcgccgctct aaagccacaa attgtccgtc acgccattcg caaactaggc ctgcactggc
     2881 gccaccgaac ggccgcgtcc aacgagcaga caccgccagc cgatccccgc gtacgttgcg
     2941 tccgtccgct gctcgaggga ggcggtggat ggagccaccc ccagttcgag aagggcggcg
     3001 gcagcggcgg cggcagcggc ggcggcagct ggagccaccc ccagttcgag aaggggcccg
     3061 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3121 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3181 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3241 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3301 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3361 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3421 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3481 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3541 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3601 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3661 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3721 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3781 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3901 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3961 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4021 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4081 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4141 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     4201 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     4261 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     4321 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     4381 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     4441 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     4501 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     4561 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     4621 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4681 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4741 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4801 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4861 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4921 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4981 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5041 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5101 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6121 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6181 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6241 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6301 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6361 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6421 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6481 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6541 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6601 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6661 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6721 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6781 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6841 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6901 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7081 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7141 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     7201 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     7261 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
