LOCUS       sequence-272760-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length mu24 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 7284)
  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 7284)
  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-mu24-2xStrep
FEATURES             Location/Qualifiers
     source          1..7284
                     /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             2174..2191
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             2949..2972
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3009..3032
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3058..3075)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3064..3288
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3334..3762
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3421..3440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3631..3652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3771..3791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3776..4105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3956..4091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4018..4037
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4153..4200
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4219..4593
                     /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    4723..4844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4760..4779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4814..4833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4893..4909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4893..4909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4906..4928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4917..4933
                     /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(4941..4947,4948..4965,4966..4971))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4986..5007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5124..5141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5295..5883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5375..5394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6054..6845,6846..6914))
                     /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     6677..6696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6915..7019)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7094..7113)
                     /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 acaatattct
      781 taccagtatt ctgtgatttg ccagaagtat accaagattg tgaatatgat gagaatagcc
      841 aactgtctat cttattctgg atatcatcca atacatatga ggtagtagag aatgtacaac
      901 aatttaatat acttggtaaa gatttaacat tctgtttatg tagagcttta agaagactgc
      961 tcctgggcat aagactgtac ccaacctgca atactgagtc taacagatat gtggttaccg
     1021 gggacagata tgtgggcaat ggtttggtgg ttagtcccac aggtaggcat attcagcgca
     1081 tagatacctg cacctatgtt ccagtgatat acagctttga acaaactgac gcacattatg
     1141 atggcatggg acctggaaaa ctgagggcct tgtatcatga gcaattattt tttaaaggaa
     1201 ttgattattc ctttgtattt tccaccctca tcagattttt atctatgaaa cagatagatg
     1261 agtgttacta ctcttttatc catactttgg aaccctgggc caggtctgct tgtaaaaaaa
     1321 attacttgaa gcttactgag tgttttaaaa atgtggccct cacaaagttg cctgtggttg
     1381 atgccacctt tcatttggaa tatatcaaat tcaatatcaa atgtttcctt gaccaatgga
     1441 atgactgccc cgacctttgg gtacttagaa gaaccatctc tgcgaattta tccaaaaaac
     1501 acctgctggc cactattgga agactcatgt ctgtgtgtga gatccagctt aggaacccct
     1561 tagcctatca tgatcaccta actatgatga aaacatttcc acaagtgaca atatctgctc
     1621 agaagaaacc caccacctgt ttaaatcaga aactggaaat agtcatgtca ggtacagagt
     1681 ccatgtggct ggtatacact cctgcttcta gtatgtttcg aattatgctc tgtgccgcct
     1741 acatcagcga catctactac aacaatactg tgcagcaggc aatttatgat gtcacccacg
     1801 ataagccaag caacggtgag atgttgatga acttgtatag aagaattgat tacctgccta
     1861 aagactcagt agaaactatt gcaagagtgt gttccacccc ctgggagaac cccctaattg
     1921 aaaggccgcc taccatgtat catgaatact gtccaatcaa aaatcttagc atcaatagtt
     1981 tcaaggtcac aatttttaat acaaatatgg tcatcaacac caaaatctca tgcaaaaaga
     2041 tcaccccccg atttagatca ttcttggata tccctaggct caccaataac tttgttgtga
     2101 aaaagtattc tgttaaagaa ccatcattta ccataagtgt gttctattct gatgatttat
     2161 gtaaaagtgg tgccatcaac atcaacatca gtggtacact aatacacttc ctctttgcta
     2221 tgggctgcct aaaatgcttc atgcccatca aaactataaa tcctgtcctg atatccaact
     2281 ggaactcaac ctttgatctt cagggactag aaaatcaaga gattgtcaga actggcaggc
     2341 atgatgtgtt ttggaccacc aacttcccat cagcggtctc taccaagcgc gggtacaaca
     2401 tttcctggtt taaggccgcc accgccaccg tctccaagat ccacggcgga gcgctgctag
     2461 atcaagtgag gggggaaatt tcacagatat tactcagcag tgatgcaatt ataaatacca
     2521 acaaaaattg catctacact acactggaaa agcgaaaccg ctttcagatt caaaccttgc
     2581 acaaaagatt cctagagtgc ttgtttgagt catgtgtggc tagcaaactt aattatcata
     2641 cagtgaaaag gctatgttct acaggcattt tcgatttcag tagacatata attagtcatt
     2701 caaaaaacaa gcatgaatgt gcactgtatg gctaccgaaa gtgtaatctt gtaccaaaaa
     2761 tattaacagg tacaaaaaag actcgcctgg acgagctggg gcggaattcc aattatcttt
     2821 cattcaataa atccccactt caccataaaa aggatgcttc ggattttaaa aaaagactgt
     2881 gccttaacag gaggagaagt aacccttgcc aaccagactc cgggcggccg ctcgagggag
     2941 gcggtggatg gagccacccc cagttcgaga agggcggcgg cagcggcggc ggcagcggcg
     3001 gcggcagctg gagccacccc cagttcgaga aggggcccgt ttaaacccgc tgatcagcct
     3061 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3121 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3181 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3241 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3301 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3361 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3421 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3481 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3541 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3601 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3661 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3721 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3781 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3841 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3901 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3961 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4021 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4081 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4141 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     4201 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     4261 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     4321 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     4381 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     4441 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     4501 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     4561 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     4621 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4681 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4741 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4801 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4861 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4921 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4981 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5041 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5101 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5161 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5221 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5281 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5341 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5401 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5461 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5521 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5581 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5641 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5701 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5761 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5821 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5881 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5941 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6001 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6061 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6121 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6181 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6241 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6301 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6361 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6421 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6481 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6541 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6601 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6661 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6721 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6781 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6841 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6901 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6961 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7021 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7081 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7141 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7201 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     7261 atgtacgggc cagatatacg cgtt
//
