LOCUS       sequence-267334-        7302 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 7302)
  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 7302)
  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..7302
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3204..3227
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3264..3287
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3313..3330)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3319..3543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3589..4017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3676..3695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3886..3907
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4026..4046)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4031..4360
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4211..4346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4273..4292
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4408..4455
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4474..4848
                     /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    4978..5099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5015..5034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5069..5088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5148..5164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5148..5164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5161..5183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5172..5188
                     /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(5196..5202,5203..5220,5221..5226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5241..5262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5379..5396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5550..6135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5630..5649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6306..7097,7098..7166))
                     /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     6929..6948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7167..7271)
                     /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 ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc catgacacaa ggtaagaggg
     1021 agatgggggg aggtctcgag ggcttctctt cacagctggg tctcttttac gccctggcct
     1081 gcaaccgcag cccacctgca cttcccgagg atgctaccct tctaatcaaa tggttggaca
     1141 cggccctggg cagggaggcc accttttacg cgtgtcgggc tatgcgtcgt cttctactcg
     1201 gcgttatccg aataaatgac tgccaggagc tgccacccgg tttaataatt ctgagtccgg
     1261 gcaccgtccc tggccccctt ggagtccaga gtctggagca tacagactgc gaaatatggt
     1321 cctctgccca ccctgaccac gctgcccacc tcccggtgcc cagggtcatc acatacaccg
     1381 actgcccggg ttccataagc acgagctcaa tgtttcgcct tatcatccgc tacttgtctc
     1441 atcaccaatt tgagcgctgc ttcgagcagt tctgccgcgt ggtcccgcgt cgcttcctag
     1501 ggacctgtaa gcaaaactct gcaaagatgc tggctagcct gaagcatctc tcgttttcaa
     1561 taatgccacc cgcatcccca gacgcacagc tacatttttt aaagtttaac atcggcagct
     1621 tcatggccac gtgggggtgg cacggagagc tggtctcgct gcgccgtgcc atcgctcaca
     1681 acgtagagcg actgcccacc gtgctgaaga acctgtcgaa acagagtaag caccaggacg
     1741 tcaaggttaa cggacgggat ctggtgggct ttcagctggc tctaaaccag ctcgtgtccc
     1801 gtctgcacgt aaaaatccaa cgcaaggacc ccggaccaaa gccatacagg gtggtcgtca
     1861 gtaccccaga ttgtacctac tatctagtgt atccgggcac accggccatc tacagactcg
     1921 tcatgtgtat ggcagtggca gactgcatcg gccactcgtg cagcggactg cacccctgcg
     1981 caaacttttt aggcacccac gagacaccgc gtctcctggc ggcgacgctt tcaagaatcc
     2041 ggtacgcgcc gaaagaccgg cgagcagcca tgaagggaaa tttgcaggcg tgcttccaac
     2101 gatacgcggc cacggacgcg cggactctgg gcagctccac agtgtcagac atgctggaac
     2161 ccacaaaaca cgtcagtttg gaaaacttca agatcaccat attcaacacc aacatggtga
     2221 ttaacactaa gataagctgc cacgttccta acaccctgca aaagactatt ttaaacatcc
     2281 ccagattgac caacaatttt gttatacgaa agtactccgt aaaggaacct tcttttacca
     2341 taagcgtgtt tttttccgac aacatgtgtc aaggcaccgc aataaacatc aacatcagtg
     2401 gggacatgct gcactttctc ttcgcaatgg gtacgctgaa atgctttctg ccaatcaggc
     2461 acatatttcc tgtatcgata gcaaattgga actccacgtt ggacctgcac ggactggaaa
     2521 accagtacat ggtgagaatg gggcgaaaaa acgtattttg gaccacaaac tttccatctg
     2581 tggtctccag caaggatggg ctaaacgtgt cctggtttaa ggccgcgaca gccacgattt
     2641 ctaaagtgta cgggcagcct cttgtggaac agattcgcca cgagctggcg cccattctca
     2701 cggaccagca cgcgcgcatc gacggaaaca aaaatagaat attctcccta cttgagcaca
     2761 gaaaccgttc ccaaatacag acgctacaca aaaggttcct ggagtgtctg gtggaatgct
     2821 gttcgtttct caggcttgac gtggcttgca ttaggcgagc cgccgcccgg ggcctgtttg
     2881 acttctcaaa gaagataatc agtcacacta aaagcaaaca cgagtgcgca gtactgggat
     2941 ataaaaagtg taacctaatc ccgaaaatct atgcccgaaa caagaagacc aggctagacg
     3001 agttgggccg caatgcaaac ttcatttcgt tcgtcgccac cacgggtcat cggttcgccg
     3061 ctctaaagcc acaaattgtc cgtcacgcca ttcgcaaact aggcctgcac tggcgccacc
     3121 gaacggccgc gtccaacgag cagacaccgc cagccgatcc ccgcgtacgt tgcgtccgtc
     3181 cgctgctcga gggaggcggt ggatggagcc acccccagtt cgagaagggc ggcggcagcg
     3241 gcggcggcag cggcggcggc agctggagcc acccccagtt cgagaagggg cccgtttaaa
     3301 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3361 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3421 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3481 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3541 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3601 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3661 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3721 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3781 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3841 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3901 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3961 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4021 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4261 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4381 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     4441 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     4501 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     4561 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     4621 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     4681 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     4741 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     4801 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     4861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4981 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5041 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5101 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6061 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     6121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6181 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6241 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6301 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6361 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6421 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6481 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6541 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6601 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6661 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6721 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6781 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6841 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6901 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6961 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7021 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7081 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7141 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7261 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
