LOCUS       sequence-267309-        7371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length ORF24 L73A/L75A with a C-terminal 3xFLAG 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-ORF24 L73A_L75A-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7371
                     /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             3291..3356
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3382..3399)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3388..3612
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3658..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3745..3764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3955..3976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4095..4115)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4100..4429
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4280..4415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4342..4361
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4477..4524
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4543..4917
                     /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    5047..5168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5084..5103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5138..5157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5217..5233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5217..5233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5230..5252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5241..5257
                     /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(5265..5271,5272..5289,5290..5295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5310..5331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5448..5465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5619..6204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5699..5718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6375..7166,7167..7235))
                     /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     6998..7017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7236..7340)
                     /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 catggcagcg ctcgagggcc
     1021 ccctactact gccaccgagc gcctccctga cgacgagtcc gcagaccacg tgttatcaag
     1081 cgacttggga atcacagctg gaaatattct gctgtctggc caccaactcg cacctgcagg
     1141 cagagctgac cttagaaggt cttgataaga tgatgcagcc cgagcccacc tttttcgcct
     1201 gcagagcgat acgcagagca ctcgcggggg aacgcctcca cccttttata catcaagaag
     1261 ggactctttt gggaaaagtg ggtcgacggt acagcggcga aggtttaata attgacggtg
     1321 gtggagtgtt tacgcgcgga cagatagaca ccgacaacta cctacctgcg gtgggatcat
     1381 gggaacttac cgatgattgt gataaaccct gcgaattcag ggagctacgc tcgctgtatc
     1441 ttcccgcgct actaacgtgc accatatgtt acaaagccat gttcaggata gtgtgcaggt
     1501 acctggagtt ctgggagttc gaacagtgtt ttcatgcgtt tctggcggtg ttgccccata
     1561 gtctacaacc cacaatctat caaaattatt ttgcactcct ggagagcctg aagcatctct
     1621 cgttttcaat aatgccaccc gcatccccag acgcacagct acatttttta aagtttaaca
     1681 tcggcagctt catggccacg tgggggtggc acggagagct ggtctcgctg cgccgtgcca
     1741 tcgctcacaa cgtagagcga ctgcccaccg tgctgaagaa cctgtcgaaa cagagtaagc
     1801 accaggacgt caaggttaac ggacgggatc tggtgggctt tcagctggct ctaaaccagc
     1861 tcgtgtcccg tctgcacgta aaaatccaac gcaaggaccc cggaccaaag ccatacaggg
     1921 tggtcgtcag taccccagat tgtacctact atctagtgta tccgggcaca ccggccatct
     1981 acagactcgt catgtgtatg gcagtggcag actgcatcgg ccactcgtgc agcggactgc
     2041 acccctgcgc aaacttttta ggcacccacg agacaccgcg tctcctggcg gcgacgcttt
     2101 caagaatccg gtacgcgccg aaagaccggc gagcagccat gaagggaaat ttgcaggcgt
     2161 gcttccaacg atacgcggcc acggacgcgc ggactctggg cagctccaca gtgtcagaca
     2221 tgctggaacc cacaaaacac gtcagtttgg aaaacttcaa gatcaccata ttcaacacca
     2281 acatggtgat taacactaag ataagctgcc acgttcctaa caccctgcaa aagactattt
     2341 taaacatccc cagattgacc aacaattttg ttatacgaaa gtactccgta aaggaacctt
     2401 cttttaccat aagcgtgttt ttttccgaca acatgtgtca aggcaccgca ataaacatca
     2461 acatcagtgg ggacatgctg cactttctct tcgcaatggg tacgctgaaa tgctttctgc
     2521 caatcaggca catatttcct gtatcgatag caaattggaa ctccacgttg gacctgcacg
     2581 gactggaaaa ccagtacatg gtgagaatgg ggcgaaaaaa cgtattttgg accacaaact
     2641 ttccatctgt ggtctccagc aaggatgggc taaacgtgtc ctggtttaag gccgcgacag
     2701 ccacgatttc taaagtgtac gggcagcctc ttgtggaaca gattcgccac gagctggcgc
     2761 ccattctcac ggaccagcac gcgcgcatcg acggaaacaa aaatagaata ttctccctac
     2821 ttgagcacag aaaccgttcc caaatacaga cgctacacaa aaggttcctg gagtgtctgg
     2881 tggaatgctg ttcgtttctc aggcttgacg tggcttgcat taggcgagcc gccgcccggg
     2941 gcctgtttga cttctcaaag aagataatca gtcacactaa aagcaaacac gagtgcgcag
     3001 tactgggata taaaaagtgt aacctaatcc cgaaaatcta tgcccgaaac aagaagacca
     3061 ggctagacga gttgggccgc aatgcaaact tcatttcgtt cgtcgccacc acgggtcatc
     3121 ggttcgccgc tctaaagcca caaattgtcc gtcacgccat tcgcaaacta ggcctgcact
     3181 ggcgccaccg aacggccgcg tccaacgagc agacaccgcc agccgatccc cgcgtacgtt
     3241 gcgtccgtcc gctggtctca gggcggccgc tcgagggagg cggtggagcc gactacaagg
     3301 accacgacgg cgactacaag gaccacgaca tcgactacaa ggacgacgac gacaaggggc
     3361 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3421 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3481 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3541 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3601 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3661 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3721 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3781 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3841 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3901 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3961 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4081 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4261 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4321 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4381 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4441 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     4501 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     4561 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     4621 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     4681 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     4741 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     4801 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     4861 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     4921 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4981 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5041 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5101 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5161 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5221 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5281 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6121 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     6181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6301 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6361 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6421 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6481 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6541 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6601 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6661 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6721 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6781 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6841 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6901 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6961 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7021 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7081 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7141 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7201 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7261 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7321 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
