LOCUS       sequence-272752-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length ORF24 L74A/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 L74A_L75A-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7374
                     /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             3057..3122
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3148..3165)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3154..3378
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3424..3852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3511..3530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3721..3742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3861..3881)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3866..4195
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4046..4181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4108..4127
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4243..4290
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4309..4683
                     /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    4813..4934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4850..4869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4904..4923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4983..4999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..4999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4996..5018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5007..5023
                     /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(5031..5037,5038..5055,5056..5061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5076..5097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5214..5231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5385..5973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5465..5484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6144..6935,6936..7004))
                     /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     6767..6786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7005..7109)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7184..7203)
                     /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 gcagcgctcg
      781 agggccccct actactgcca ccgagcgcct ccctgacgac gagtccgcag accacgtgtt
      841 atcaagcgac ttgggaatca cagctggaaa tattctgctg tctggccacc aactcgcacc
      901 tgcaggcaga gctgacctta gaaggtcttg ataagatgat gcagcccgag cccacctttt
      961 tcgcctgcag agcgatacgc agactagccg cgggggaacg cctccaccct tttatacatc
     1021 aagaagggac tcttttggga aaagtgggtc gacggtacag cggcgaaggt ttaataattg
     1081 acggtggtgg agtgtttacg cgcggacaga tagacaccga caactaccta cctgcggtgg
     1141 gatcatggga acttaccgat gattgtgata aaccctgcga attcagggag ctacgctcgc
     1201 tgtatcttcc cgcgctacta acgtgcacca tatgttacaa agccatgttc aggatagtgt
     1261 gcaggtacct ggagttctgg gagttcgaac agtgttttca tgcgtttctg gcggtgttgc
     1321 cccatagtct acaacccaca atctatcaaa attattttgc actcctggag agcctgaagc
     1381 atctctcgtt ttcaataatg ccacccgcat ccccagacgc acagctacat tttttaaagt
     1441 ttaacatcgg cagcttcatg gccacgtggg ggtggcacgg agagctggtc tcgctgcgcc
     1501 gtgccatcgc tcacaacgta gagcgactgc ccaccgtgct gaagaacctg tcgaaacaga
     1561 gtaagcacca ggacgtcaag gttaacggac gggatctggt gggctttcag ctggctctaa
     1621 accagctcgt gtcccgtctg cacgtaaaaa tccaacgcaa ggaccccgga ccaaagccat
     1681 acagggtggt cgtcagtacc ccagattgta cctactatct agtgtatccg ggcacaccgg
     1741 ccatctacag actcgtcatg tgtatggcag tggcagactg catcggccac tcgtgcagcg
     1801 gactgcaccc ctgcgcaaac tttttaggca cccacgagac accgcgtctc ctggcggcga
     1861 cgctttcaag aatccggtac gcgccgaaag accggcgagc agccatgaag ggaaatttgc
     1921 aggcgtgctt ccaacgatac gcggccacgg acgcgcggac tctgggcagc tccacagtgt
     1981 cagacatgct ggaacccaca aaacacgtca gtttggaaaa cttcaagatc accatattca
     2041 acaccaacat ggtgattaac actaagataa gctgccacgt tcctaacacc ctgcaaaaga
     2101 ctattttaaa catccccaga ttgaccaaca attttgttat acgaaagtac tccgtaaagg
     2161 aaccttcttt taccataagc gtgttttttt ccgacaacat gtgtcaaggc accgcaataa
     2221 acatcaacat cagtggggac atgctgcact ttctcttcgc aatgggtacg ctgaaatgct
     2281 ttctgccaat caggcacata tttcctgtat cgatagcaaa ttggaactcc acgttggacc
     2341 tgcacggact ggaaaaccag tacatggtga gaatggggcg aaaaaacgta ttttggacca
     2401 caaactttcc atctgtggtc tccagcaagg atgggctaaa cgtgtcctgg tttaaggccg
     2461 cgacagccac gatttctaaa gtgtacgggc agcctcttgt ggaacagatt cgccacgagc
     2521 tggcgcccat tctcacggac cagcacgcgc gcatcgacgg aaacaaaaat agaatattct
     2581 ccctacttga gcacagaaac cgttcccaaa tacagacgct acacaaaagg ttcctggagt
     2641 gtctggtgga atgctgttcg tttctcaggc ttgacgtggc ttgcattagg cgagccgccg
     2701 cccggggcct gtttgacttc tcaaagaaga taatcagtca cactaaaagc aaacacgagt
     2761 gcgcagtact gggatataaa aagtgtaacc taatcccgaa aatctatgcc cgaaacaaga
     2821 agaccaggct agacgagttg ggccgcaatg caaacttcat ttcgttcgtc gccaccacgg
     2881 gtcatcggtt cgccgctcta aagccacaaa ttgtccgtca cgccattcgc aaactaggcc
     2941 tgcactggcg ccaccgaacg gccgcgtcca acgagcagac accgccagcc gatccccgcg
     3001 tacgttgcgt ccgtccgctg gtctcagggc ggccgctcga gggaggcggt ggagccgact
     3061 acaaggacca cgacggcgac tacaaggacc acgacatcga ctacaaggac gacgacgaca
     3121 aggggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3181 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3241 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3301 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3361 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3421 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3841 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3901 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3961 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4021 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4201 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     4261 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     4321 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     4381 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     4441 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     4501 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     4561 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     4621 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     4681 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4801 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4861 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4921 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4981 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5041 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5101 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7141 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     7201 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7261 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7321 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
