LOCUS       sequence-292094-        7691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  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 7691)
  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-ORF8-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..7691
                     /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             3590..3613
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3650..3673
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3699..3716)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3705..3929
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3975..4403
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4062..4081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4272..4293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4412..4432)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4417..4746
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4597..4732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4659..4678
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4794..4841
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4860..5234
                     /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    5364..5485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5401..5420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5455..5474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5534..5550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5534..5550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5547..5569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5558..5574
                     /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(5582..5588,5589..5606,5607..5612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5627..5648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5765..5782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5936..6524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6016..6035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6695..7486,7487..7555))
                     /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     7318..7337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7556..7660)
                     /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 attcatgact cccaggtcta gattggccac cctggggact gtcatcctgt tggtctgctt
     1081 ttgcgcaggc gcggcgcact cgaggggtga cacctttcag acgtccagtt cccccacacc
     1141 cccaggatct tcctctaagg cccccaccaa acctggtgag gaagcatctg gtcctaagag
     1201 tgtggacttt taccagttca gagtgtgtag tgcatcgatc accggggagc tttttcggtt
     1261 caacctggag cagacgtgcc cagacaccaa agacaagtac caccaagaag gaattttact
     1321 ggtgtacaaa aaaaacatag tgcctcatat ctttaaggtg cggcgctata ggaaaattgc
     1381 cacctctgtc acggtctaca ggggcttgac agagtccgcc atcaccaaca agtatgaact
     1441 cccgagaccc gtgccactct atgagataag ccacatggac agcacctatc agtgctttag
     1501 ttccatgaag gtaaatgtca acggggtaga aaacacattt actgacagag acgatgttaa
     1561 caccacagta ttcctccaac cagtagaggg gcttacggat aacattcaaa ggtactttag
     1621 ccagccggtc atctacgcgg aacccggctg gtttcccggc atatacagag ttaggaccac
     1681 cgtcaattgc gagatagtgg acatgatagc caggtctgct gaaccataca attactttgt
     1741 cacgtcactg ggtgacacgg tggaagtctc ccctttttgc tataacgaat cctcatgcag
     1801 cacaaccccc agcaacaaaa atggccttag cgtccaagta gttctcaacc acactgtggt
     1861 cacgtactct gacagaggaa ccagtcccac tccccaaaac aggatctttg tggaaacggg
     1921 agcgtacacg ctttcgtggg cctccgagag caagaccacg gccgtgtgtc cgctggcact
     1981 gtggaaaacc ttcccgcgct ccatccagac tacccacgag gacagcttcc actttgtggc
     2041 caacgagatc acggccacct tcacggctcc tctaacgcca gtggccaact ttaccgacac
     2101 gtactcttgt ctgacctcgg atatcaacac cacgctaaac gccagcaagg ccaaactggc
     2161 gagcactcac gtccctaacg ggacggtcca gtacttccac acaacaggcg gactctattt
     2221 ggtctggcag cccatgtccg cgattaacct gactcacgct cagggcgaca gcgggaaccc
     2281 cacgtcatcg ccgcccccct ccgcatcccc catgaccacc tctgccagcc gcagaaagag
     2341 acggtcagcc agtaccgctg ctgccggcgg cggggggtcc acggacaacc tgtcttacac
     2401 gcagctgcag tttgcctacg acaaactgcg ggatggcatt aatcaggtgt tagaagaact
     2461 ctccagggca tggtgtcgcg agcaggtcag ggacaaccta atgtggtacg agctcagtaa
     2521 aatcaacccc accagcgtta tgacagccat ctacggtcga cctgtatccg ccaagttcgt
     2581 aggagacgcc atttccgtga ccgagtgcat taacgtggac cagagctccg taaacatcca
     2641 caagagcctc agaaccaata gtaaggacgt gtgttacgcg cgccccctgg tgacgtttaa
     2701 gtttttgaac agttccaacc tattcaccgg ccagctgggc gcgcgcaatg agataatact
     2761 gaccaacaac caggtggaaa cctgcaaaga cacctgcgaa cactacttca tcacccgcaa
     2821 cgagactctg gtgtataagg actacgcgta cctgcgcact ataaacacca ctgacatatc
     2881 caccctgaac acttttatcg ccctgaatct atcctttatt caaaacatag acttcaaggc
     2941 catcgagctg tacagcagtg cagagaaacg actcgcgagt agcgtgtttg acctggagac
     3001 gatgttcagg gagtacaact actacacaca tcgtctcgcg ggtttgcgcg aggatctgga
     3061 caacaccata gatatgaaca aggagcgctt cgtaagggac ttgtcggaga tagtggcgga
     3121 cctgggtggc atcggaaaaa cggtggtgaa cgtggccagc agcgtggtca ctctatgtgg
     3181 ctcattggtt accggattca taaattttat taaacacccc ctaggtggca tgctgatgat
     3241 cattatcgtt atagcaatca tcctgatcat ttttatgctc agtcgccgca ccaataccat
     3301 agcccaggcg ccggtgaaga tgatctaccc cgacgtagat cgcagggcac ctcctagcgg
     3361 cggagcccca acacgggagg aaatcaaaaa catcctgctg ggaatgcacc agctacaaca
     3421 agaggagagg cagaaggcgg atgatatgaa aaaaagtaca ccctcggtgt ttcagcgtac
     3481 cgcaaacggc cttcgtcagc gtctgagagg atataaacct ctgactcaat cgctagacat
     3541 cagtccggaa acgggggagg gagcgcggcc gctcgaggga ggcggtggat ggagccaccc
     3601 ccagttcgag aagggcggcg gcagcggcgg cggcagcggc ggcggcagct ggagccaccc
     3661 ccagttcgag aaggggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3721 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3781 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3841 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3901 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3961 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4021 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4081 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4141 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4201 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4261 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4321 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4381 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4441 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4501 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4561 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4621 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4681 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4741 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     4801 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     4861 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg
     4921 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg
     4981 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca
     5041 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg
     5101 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc
     5161 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg
     5221 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     5281 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5341 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5401 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5461 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5521 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5581 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5641 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5701 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5761 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5821 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5881 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5941 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6001 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6061 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6121 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6181 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6241 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6301 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6361 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6421 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6481 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6541 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6601 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6661 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6721 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6781 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6841 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6901 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6961 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7021 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7081 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7141 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7201 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7261 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7321 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7381 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7441 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7501 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7561 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7621 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7681 cacctgacgt c
//
