LOCUS       sequence-272748-        5724 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses truncated ORF24 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 5724)
  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 5724)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 1-201-2xStrep
FEATURES             Location/Qualifiers
     source          1..5724
                     /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             1389..1412
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1449..1472
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1498..1515)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1504..1728
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1774..2202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1861..1880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2071..2092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2211..2231)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2216..2545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2396..2531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2458..2477
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2593..2640
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2659..3033
                     /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    3163..3284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3200..3219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3254..3273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3333..3349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3333..3349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3346..3368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3357..3373
                     /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(3381..3387,3388..3405,3406..3411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3426..3447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3564..3581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3735..4323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3815..3834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4494..5285,5286..5354))
                     /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     5117..5136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5355..5459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5534..5553)
                     /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 ctcgagggag
     1381 gcggtggatg gagccacccc cagttcgaga agggcggcgg cagcggcggc ggcagcggcg
     1441 gcggcagctg gagccacccc cagttcgaga aggggcccgt ttaaacccgc tgatcagcct
     1501 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1561 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1621 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1681 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     1741 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     1801 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1861 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1921 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1981 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2041 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2101 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2161 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2221 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2281 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2341 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2401 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2461 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2521 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     2581 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     2641 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     2701 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     2761 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     2821 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     2881 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     2941 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3001 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     3061 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     3121 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     3181 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3241 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     3301 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3361 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3421 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3541 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3601 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3661 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3721 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3901 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4141 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5341 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5401 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5461 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     5521 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     5581 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     5641 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     5701 atgtacgggc cagatatacg cgtt
//
