LOCUS       sequence-272771-        7371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length BcRF1 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 7371)
  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 7371)
  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-BcRF1-2xStrep
FEATURES             Location/Qualifiers
     source          1..7371
                     /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             3036..3059
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3096..3119
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3145..3162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3151..3375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3421..3849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3508..3527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3718..3739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3858..3878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3863..4192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4043..4178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4105..4124
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4240..4287
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4306..4680
                     /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    4810..4931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4847..4866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4901..4920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4980..4996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4980..4996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4993..5015)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5004..5020
                     /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(5028..5034,5035..5052,5053..5058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5073..5094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5211..5228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5382..5970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5462..5481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6141..6932,6933..7001))
                     /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     6764..6783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7002..7106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7181..7200)
                     /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 tcatctgaag caggttacca
     1321 ggatcccccc ctgtccgccc ttcagcgggc gggaggccag actcaagttc cacttcttct
     1381 cctggagcac attcatgctg tcatggccaa acaatgccac actccgggag atcaggacga
     1441 gggccgccac caacctcacc caccacccac atctagtgga cactctgtac cacgcctctc
     1501 cacagacccc atttctgaca cgcagcggtg ctctataccg cttcgtcacc tgttgcaact
     1561 gcaccctgcc caatatctcc atccagcagt gcaaggccgg ggacagaccg ggggacctgg
     1621 agatcattct acagagtaac ggcggaggga ggcctgcgag cttccagttc ccctcctccc
     1681 caaccggcgc cctattgcga tgcatagttg ctgcggccct gctgccagag gtgtccgtgg
     1741 ggcaccagga gctgtctccg ctcatgtcca gaagccatgg agggcagacg gatgtcaggt
     1801 cgggcccgga cccggcccgg aggctggtgg ccctcctgcg aagggaagat ggggcaccta
     1861 aggacccccc tctgggaccg tttggacacc cccgggggcc cggcccggcc aagagcgaag
     1921 acgaggagtc tgagcgtcga gacgcccctc cacccccgct cgatttcagc ttccaagctt
     1981 cccggttggt gcccgtgggg cctgggtttc gcctgctcgt gttcaacacc aatcgggtga
     2041 tcaacactaa attggtgtgc tcagagcccc tggtgaagat gcgagtttgc aatgtccccc
     2101 gcctcatcaa caactttgta gcccgcaagt acgtggtgaa agagacggcg ttcaccgtca
     2161 gtctattctt tacggacggg gtgggggcca acttagccat caatgtcaat atcagtggca
     2221 cctatctgag cttcctattg gccatgacgt cactgcggtg cttcctgcct gtagaggcta
     2281 tttatcccgc ggccgtgtca aactggaact cgactctaga tctccatggg ctggaaaatc
     2341 agagcctagt cagagagaac cgaagcgggg tcttttggac taccaacttt ccctcggtgg
     2401 tgtcctgcca ggacggtctc aacgtgtcct ggtttaaggc cgcaactgcc accatatctc
     2461 gagtgcacgg gcggacattg gagcagcacc tgatccgtga aatcaccccc atcgtgacgc
     2521 atcgagaggc aaaaatctcc cggattaaaa accggctctt taccctgcta gagctacgca
     2581 atcggagtca gattcaagtg ctgcacaagc gtttcctgga aggcctgcta gactgcgcct
     2641 ccctcctgcg cctggatccc agctgtatca accgaatcgc ctccgagggc ctgtttgatt
     2701 tctccaagag aagcatcgcc cactccaaaa accgacacga gtgcgcgctt ctgggtcaca
     2761 gacattcggc gaacgtgaca aagcttgtgg taaacgagcg caagacccgc ctggacatac
     2821 tgggccgtaa cgctaacttt ttaacgaggt gtaagcatca ggttaatcta agacagtcac
     2881 ctattttcct gaccctcctg aggcacatcc gccgacgtct gggcctgggc cgtgcttccg
     2941 taaaacgaga gattaccctt ctcctggccc acctgcgcaa aaagacagcc cccatccact
     3001 gccgtgatgc tcaagtgctc gagggaggcg gtggatggag ccacccccag ttcgagaagg
     3061 gcggcggcag cggcggcggc agcggcggcg gcagctggag ccacccccag ttcgagaagg
     3121 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3181 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3241 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3301 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3361 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3421 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3481 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3541 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3601 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3661 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3721 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3781 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3841 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4021 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4141 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4201 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     4261 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     4321 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     4381 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     4441 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     4501 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     4561 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     4621 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     4681 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4741 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4801 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4861 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4921 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4981 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5041 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5101 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5161 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6181 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6241 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6301 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6361 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6421 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6481 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6541 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6601 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6661 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6721 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6781 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6841 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6901 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6961 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7081 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7141 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     7201 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     7261 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     7321 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
