LOCUS       sequence-267319-        7368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length BcRF1 3L_A 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 7368)
  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 7368)
  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 3L_A-2xStrep
FEATURES             Location/Qualifiers
     source          1..7368
                     /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             3270..3293
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3330..3353
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3379..3396)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3385..3609
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3655..4083
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3742..3761)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3952..3973
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4092..4112)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4097..4426
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4277..4412
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4339..4358
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4474..4521
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4540..4914
                     /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    5044..5165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5081..5100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5135..5154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5214..5230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5214..5230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5227..5249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5238..5254
                     /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(5262..5268,5269..5286,5287..5292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5307..5328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5445..5462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5616..6201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5696..5715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6372..7163,7164..7232))
                     /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     6995..7014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7233..7337)
                     /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 gccgcggcag
     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 tggctcatct gaagcaggtt accaggatcc
     1561 ccccctgtcc gcccttcagc gggcgggagg ccagactcaa gttccacttc ttctcctgga
     1621 gcacattcat gctgtcatgg ccaaacaatg ccacactccg ggagatcagg acgagggccg
     1681 ccaccaacct cacccaccac ccacatctag tggacactct gtaccacgcc tctccacaga
     1741 ccccatttct gacacgcagc ggtgctctat accgcttcgt cacctgttgc aactgcaccc
     1801 tgcccaatat ctccatccag cagtgcaagg ccggggacag accgggggac ctggagatca
     1861 ttctacagag taacggcgga gggaggcctg cgagcttcca gttcccctcc tccccaaccg
     1921 gcgccctatt gcgatgcata gttgctgcgg ccctgctgcc agaggtgtcc gtggggcacc
     1981 aggagctgtc tccgctcatg tccagaagcc atggagggca gacggatgtc aggtcgggcc
     2041 cggacccggc ccggaggctg gtggccctcc tgcgaaggga agatggggca cctaaggacc
     2101 cccctctggg accgtttgga cacccccggg ggcccggccc ggccaagagc gaagacgagg
     2161 agtctgagcg tcgagacgcc cctccacccc cgctcgattt cagcttccaa gcttcccggt
     2221 tggtgcccgt ggggcctggg tttcgcctgc tcgtgttcaa caccaatcgg gtgatcaaca
     2281 ctaaattggt gtgctcagag cccctggtga agatgcgagt ttgcaatgtc ccccgcctca
     2341 tcaacaactt tgtagcccgc aagtacgtgg tgaaagagac ggcgttcacc gtcagtctat
     2401 tctttacgga cggggtgggg gccaacttag ccatcaatgt caatatcagt ggcacctatc
     2461 tgagcttcct attggccatg acgtcactgc ggtgcttcct gcctgtagag gctatttatc
     2521 ccgcggccgt gtcaaactgg aactcgactc tagatctcca tgggctggaa aatcagagcc
     2581 tagtcagaga gaaccgaagc ggggtctttt ggactaccaa ctttccctcg gtggtgtcct
     2641 gccaggacgg tctcaacgtg tcctggttta aggccgcaac tgccaccata tctcgagtgc
     2701 acgggcggac attggagcag cacctgatcc gtgaaatcac ccccatcgtg acgcatcgag
     2761 aggcaaaaat ctcccggatt aaaaaccggc tctttaccct gctagagcta cgcaatcgga
     2821 gtcagattca agtgctgcac aagcgtttcc tggaaggcct gctagactgc gcctccctcc
     2881 tgcgcctgga tcccagctgt atcaaccgaa tcgcctccga gggcctgttt gatttctcca
     2941 agagaagcat cgcccactcc aaaaaccgac acgagtgcgc gcttctgggt cacagacatt
     3001 cggcgaacgt gacaaagctt gtggtaaacg agcgcaagac ccgcctggac atactgggcc
     3061 gtaacgctaa ctttttaacg aggtgtaagc atcaggttaa tctaagacag tcacctattt
     3121 tcctgaccct cctgaggcac atccgccgac gtctgggcct gggccgtgct tccgtaaaac
     3181 gagagattac ccttctcctg gcccacctgc gcaaaaagac agcccccatc cactgccgtg
     3241 atgctcaagt gctcgaggga ggcggtggat ggagccaccc ccagttcgag aagggcggcg
     3301 gcagcggcgg cggcagcggc ggcggcagct ggagccaccc ccagttcgag aaggggcccg
     3361 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3421 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3481 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3541 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3601 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3661 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3721 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3781 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3841 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3901 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3961 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4021 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4081 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4141 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4261 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4321 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4381 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4441 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     4501 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     4561 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     4621 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     4681 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     4741 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     4801 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     4861 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     4921 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4981 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5041 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5101 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5161 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5221 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5281 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5341 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5401 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6121 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     6181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6661 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6961 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7321 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
