LOCUS       sequence-272838-        7374 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 7374)
  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 7374)
  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..7374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     255..279
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2677..2742
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2768..2785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2774..2998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3044..3472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3131..3150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3341..3362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3481..3501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3486..3815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3666..3801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3728..3747
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3863..3910
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3929..4303
                     /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    4433..4554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4470..4489)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4524..4543
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4603..4619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4603..4619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4616..4638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4627..4643
                     /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(4651..4657,4658..4675,4676..4681))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4696..4717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4834..4851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5005..5593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5085..5104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5764..6555,6556..6624))
                     /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     6387..6406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6625..6729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6804..6823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6995..7374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
      241 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      301 tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct agcgtttaaa
      361 cttaagcttg gtaccgagct cggatccatg gcagcgctcg agggccccct actactgcca
      421 ccgagcgcct ccctgacgac gagtccgcag accacgtgtt atcaagcgac ttgggaatca
      481 cagctggaaa tattctgctg tctggccacc aactcgcacc tgcaggcaga gctgacctta
      541 gaaggtcttg ataagatgat gcagcccgag cccacctttt tcgcctgcag agcgatacgc
      601 agagcactcg cgggggaacg cctccaccct tttatacatc aagaagggac tcttttggga
      661 aaagtgggtc gacggtacag cggcgaaggt ttaataattg acggtggtgg agtgtttacg
      721 cgcggacaga tagacaccga caactaccta cctgcggtgg gatcatggga acttaccgat
      781 gattgtgata aaccctgcga attcagggag ctacgctcgc tgtatcttcc cgcgctacta
      841 acgtgcacca tatgttacaa agccatgttc aggatagtgt gcaggtacct ggagttctgg
      901 gagttcgaac agtgttttca tgcgtttctg gcggtgttgc cccatagtct acaacccaca
      961 atctatcaaa attattttgc actcctggag agcctgaagc atctctcgtt ttcaataatg
     1021 ccacccgcat ccccagacgc acagctacat tttttaaagt ttaacatcgg cagcttcatg
     1081 gccacgtggg ggtggcacgg agagctggtc tcgctgcgcc gtgccatcgc tcacaacgta
     1141 gagcgactgc ccaccgtgct gaagaacctg tcgaaacaga gtaagcacca ggacgtcaag
     1201 gttaacggac gggatctggt gggctttcag ctggctctaa accagctcgt gtcccgtctg
     1261 cacgtaaaaa tccaacgcaa ggaccccgga ccaaagccat acagggtggt cgtcagtacc
     1321 ccagattgta cctactatct agtgtatccg ggcacaccgg ccatctacag actcgtcatg
     1381 tgtatggcag tggcagactg catcggccac tcgtgcagcg gactgcaccc ctgcgcaaac
     1441 tttttaggca cccacgagac accgcgtctc ctggcggcga cgctttcaag aatccggtac
     1501 gcgccgaaag accggcgagc agccatgaag ggaaatttgc aggcgtgctt ccaacgatac
     1561 gcggccacgg acgcgcggac tctgggcagc tccacagtgt cagacatgct ggaacccaca
     1621 aaacacgtca gtttggaaaa cttcaagatc accatattca acaccaacat ggtgattaac
     1681 actaagataa gctgccacgt tcctaacacc ctgcaaaaga ctattttaaa catccccaga
     1741 ttgaccaaca attttgttat acgaaagtac tccgtaaagg aaccttcttt taccataagc
     1801 gtgttttttt ccgacaacat gtgtcaaggc accgcaataa acatcaacat cagtggggac
     1861 atgctgcact ttctcttcgc aatgggtacg ctgaaatgct ttctgccaat caggcacata
     1921 tttcctgtat cgatagcaaa ttggaactcc acgttggacc tgcacggact ggaaaaccag
     1981 tacatggtga gaatggggcg aaaaaacgta ttttggacca caaactttcc atctgtggtc
     2041 tccagcaagg atgggctaaa cgtgtcctgg tttaaggccg cgacagccac gatttctaaa
     2101 gtgtacgggc agcctcttgt ggaacagatt cgccacgagc tggcgcccat tctcacggac
     2161 cagcacgcgc gcatcgacgg aaacaaaaat agaatattct ccctacttga gcacagaaac
     2221 cgttcccaaa tacagacgct acacaaaagg ttcctggagt gtctggtgga atgctgttcg
     2281 tttctcaggc ttgacgtggc ttgcattagg cgagccgccg cccggggcct gtttgacttc
     2341 tcaaagaaga taatcagtca cactaaaagc aaacacgagt gcgcagtact gggatataaa
     2401 aagtgtaacc taatcccgaa aatctatgcc cgaaacaaga agaccaggct agacgagttg
     2461 ggccgcaatg caaacttcat ttcgttcgtc gccaccacgg gtcatcggtt cgccgctcta
     2521 aagccacaaa ttgtccgtca cgccattcgc aaactaggcc tgcactggcg ccaccgaacg
     2581 gccgcgtcca acgagcagac accgccagcc gatccccgcg tacgttgcgt ccgtccgctg
     2641 gtctcagggc ggccgctcga gggaggcggt ggagccgact acaaggacca cgacggcgac
     2701 tacaaggacc acgacatcga ctacaaggac gacgacgaca aggggcccgt ttaaacccgc
     2761 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2821 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2881 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2941 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3001 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3061 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3121 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3181 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3241 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3301 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3361 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3421 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3481 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3541 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3661 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3721 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3781 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3841 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3901 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
     3961 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
     4021 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     4081 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     4141 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     4201 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     4261 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
     4321 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4381 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4441 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4501 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4561 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4621 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4681 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4741 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4801 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4861 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4921 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4981 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5041 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5101 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5161 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5221 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5281 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5341 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5401 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5461 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5521 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5581 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5641 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5701 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5761 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5821 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5881 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5941 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6001 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6061 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6121 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6181 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6241 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6301 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6361 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6421 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6481 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6541 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6601 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6661 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6721 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6781 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6841 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     6901 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     6961 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     7021 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     7081 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     7141 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     7201 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     7261 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7321 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catg
//
