LOCUS       sequence-272726-        5718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated ORF24 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 5718)
  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 5718)
  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 1-201-3xFLAG
FEATURES             Location/Qualifiers
     source          1..5718
                     /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             1401..1466
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1492..1509)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1498..1722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1768..2196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1855..1874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2065..2086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2205..2225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2210..2539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2390..2525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2452..2471
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2587..2634
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2653..3027
                     /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    3157..3278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3194..3213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3248..3267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3327..3343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3327..3343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3340..3362)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3351..3367
                     /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(3375..3381,3382..3399,3400..3405))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3420..3441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3558..3575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3729..4317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3809..3828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4488..5279,5280..5348))
                     /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     5111..5130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5349..5453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5528..5547)
                     /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 agactactcc tgggggaacg 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 gggcggccgc
     1381 tcgagggagg cggtggagcc gactacaagg accacgacgg cgactacaag gaccacgaca
     1441 tcgactacaa ggacgacgac gacaaggggc ccgtttaaac ccgctgatca gcctcgactg
     1501 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1561 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1621 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1681 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1741 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     1801 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1861 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1921 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1981 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2041 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2101 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2161 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2221 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2281 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2341 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2401 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2461 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     2521 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     2581 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     2641 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     2701 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     2761 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     2821 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     2881 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     2941 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     3001 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     3061 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3121 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3181 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3241 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3301 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3361 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3421 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3481 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3541 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3601 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3661 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3721 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3781 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3841 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3901 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3961 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4021 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4081 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4141 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4201 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4261 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4321 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4381 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4441 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4501 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4561 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4621 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4681 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4741 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4801 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4861 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4921 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4981 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5041 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5101 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5161 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5221 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5281 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5341 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5401 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5461 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     5521 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     5581 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     5641 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     5701 gggccagata tacgcgtt
//
