LOCUS       sequence-272758-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length ORF24 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 7377)
  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 7377)
  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 3L_A-2xStrep
FEATURES             Location/Qualifiers
     source          1..7377
                     /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             3042..3065
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3102..3125
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3151..3168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3157..3381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3427..3855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3514..3533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3724..3745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3864..3884)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3869..4198
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4049..4184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4111..4130
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4246..4293
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4312..4686
                     /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    4816..4937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4853..4872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4907..4926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4986..5002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4986..5002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4999..5021)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5010..5026
                     /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(5034..5040,5041..5058,5059..5064))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5079..5100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5217..5234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5388..5976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5468..5487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6147..6938,6939..7007))
                     /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     6770..6789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7008..7112)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7187..7206)
                     /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 agagcagccg 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 gtcctcgagg gaggcggtgg atggagccac ccccagttcg
     3061 agaagggcgg cggcagcggc ggcggcagcg gcggcggcag ctggagccac ccccagttcg
     3121 agaaggggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3181 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3241 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3301 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3361 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3421 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3481 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3541 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3601 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3661 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3721 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3781 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3841 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3901 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3961 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4021 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4081 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4141 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4201 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
     4261 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
     4321 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     4381 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     4441 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     4501 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     4561 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     4621 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     4681 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4741 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4801 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4861 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4921 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4981 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5041 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5101 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5161 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5221 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5281 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5821 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6121 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6181 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6241 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6301 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6361 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6421 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6481 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6541 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6601 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6661 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6721 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6781 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6841 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6901 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6961 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7021 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7081 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7141 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     7201 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7261 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7321 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
