LOCUS       sequence-267411-        6806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  AUTHORS   Davis ZH, Verschueren E, Jang GM, Kleffman K, Johnson JR, Park J, 
            Von Dollen J, Maher MC, Johnson T, Newton W, Jager S, Shales M, 
            Horner J, Hernandez RD, Krogan NJ, Glaunsinger BA
  TITLE     Global mapping of herpesvirus-host protein complexes reveals a 
            transcription strategy for late genes.
  JOURNAL   Mol Cell. 2015 Jan 22;57(2):349-60. doi: 
            10.1016/j.molcel.2014.11.026. Epub 2014 Dec 24.
  PUBMED    25544563
REFERENCE   2  (bases 1 to 6806)
  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"; doi: 
            "10.1016/j.molcel.2014.11.026"; journalName: "Mol Cell"; date: 
            "2015-01-22- 22"; volume: "57"; issue: "2"; pages: "349-60"
COMMENT     Sequence Label: pCDNA4.TO-ORF4-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..6806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2705..2728
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2765..2788
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2814..2831)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2820..3044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3090..3518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3177..3196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3387..3408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3527..3547)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3532..3861
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3712..3847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3774..3793
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3909..3956
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3975..4349
                     /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    4479..4600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4516..4535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4570..4589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4649..4665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4649..4665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4662..4684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4673..4689
                     /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(4697..4703,4704..4721,4722..4727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4742..4763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4880..4897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5051..5639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5131..5150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5810..6601,6602..6670))
                     /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     6433..6452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6671..6775)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attcatggcc tttttaagac aaacactgtg gattttatgg acatttacca tggtcattgg
     1081 ccaggacaat gaaaagtgtt cccaaaaaac cttaattgga tatagactta aaatgtctcg
     1141 tgacggtgac attgcagttg gagaaacagt ggaattacgt tgtagatctg gatacactac
     1201 ttatgcccgc aatataacag caacatgttt acaaggtggg acgtggtctg aaccaacggc
     1261 aacatgtaac aaaaagtcct gtccaaaccc aggtgaaata caaaatggaa aggttatatt
     1321 tcatggtgga caagatgcct taaaatatgg ggcaaacatt tcatatgttt gtaatgaagg
     1381 atattttttg gttggtcgag aatacgtgcg atattgtatg attggagcat ctggccaaat
     1441 ggcgtggtca tcttctcctc ctttttgtga aaaagaaaag tgtcacagac cgaaaatcga
     1501 aaatggagat tttaagcctg ataaagatta ttatgagtat aatgatgcag ttcattttga
     1561 atgtaatgaa ggatatactc tagttggacc acattccatt gcatgtgcag ttaataacac
     1621 gtggacatct aacatgccaa cctgtgaact cgcaggctgt aaatttccat cggtgactca
     1681 tggttatcca atccaaggtt tttctcttac ttataaacat aagcaaagtg ttacttttgc
     1741 atgcaatgat ggatttgttc tcagaggatc ccccacaatt acgtgtaacg ttactgaatg
     1801 ggacccacca cttcctaagt gtgttttgga agatatagat gatccaaaca attcaaatcc
     1861 tggacgtttg catccaacac ccaatgaaaa accaaatggt aatgtctttc aacgctcaaa
     1921 ctatacagaa cctccaacaa agcctgaaga cacccataca gcagctactt gtgataccaa
     1981 ctgtgaacag ccacctaaaa tcctgccaac atccgaaggt tttaatgaga ctaccacatc
     2041 taatacaatt acaaaacaat tagaggatga gaaaactaca tcccagccaa atacacatat
     2101 tacatctgcc ttaacatcca tgaaagcgaa aggtaacttt accaacaaga ccaataactc
     2161 tactgatcta catatagcat ctacacccac ttcccaagat gatgctacgc cttcaatacc
     2221 tagtgtacag acacccaatt ataatactaa cgcaccgaca cgtacactaa cgtctctcca
     2281 tattgaagaa ggcccatcca attctactac ttcagaaaag gccactgcct ctactctctc
     2341 acacaactca cacaaaaatg acaccggagg catatacaca acattaaaca aaacaacaca
     2401 gttgccatcc actaataaac ctacaaacag tcaagccaag agttccacta agccacgcgt
     2461 tgagacacac aataaaacaa ccagtaatcc tgccatttct ttaacagatt ctgcagatgt
     2521 gcctcagaga ccgcgagaac caacactccc tcccattttc aggccaccgg cgtctaaaaa
     2581 tcgctatctg gaaaagcaac tagttattgg actactaacc gctgtcgccc taacgtgtgg
     2641 actgattacc ttatttcact atctgttctt tcgttcgggg cggccgctcg agggaggcgg
     2701 tggatggagc cacccccagt tcgagaaggg cggcggcagc ggcggcggca gcggcggcgg
     2761 cagctggagc cacccccagt tcgagaaggg gcccgtttaa acccgctgat cagcctcgac
     2821 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2881 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2941 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3001 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3061 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3121 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3181 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3241 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3301 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3361 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3421 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3481 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3541 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3661 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3721 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3781 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3841 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3901 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3961 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     4021 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     4081 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     4141 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     4201 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     4261 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     4321 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     4381 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4441 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4621 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4681 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4741 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5461 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6781 ttccccgaaa agtgccacct gacgtc
//
