LOCUS       sequence-267649-        8964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF75.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8964)
  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 8964)
  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-ORF75-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..8964
                     /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             4863..4886
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4923..4946
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4972..4989)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4978..5202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5248..5676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5335..5354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5545..5566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5685..5705)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5690..6019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5870..6005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5932..5951
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6067..6114
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6133..6507
                     /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    6637..6758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6674..6693)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6728..6747
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6807..6823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6807..6823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6820..6842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6831..6847
                     /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(6855..6861,6862..6879,6880..6885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6900..6921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7038..7055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7209..7797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7289..7308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7968..8759,8760..8828))
                     /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     8591..8610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8829..8933)
                     /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 catgtttgtc agcttttata
     1021 caagccgtac gggcacactc actctcgtac ccggtggcac cggcggctac tatctgctat
     1081 ggataacttt ccgaagacct cccacttcga gggaggagcg agagagacgg gatgtggaaa
     1141 tacagacggt gctcgctgtg ctgtcaccgc tccttggata tccccatgtc atcaggcggt
     1201 ctccgccacg ggggagcgag cgtgttgtat cctttggcta cgggccaaac ataaaccaca
     1261 ggcccacaac actgtcaaca gagcttggag ttttgctgcg agagctagga ctgcaggagt
     1321 gggctagagt ggaagtgggc aggcacctgg tgtccaaaat cacacagacc ctgctagaac
     1381 cacacccacc tcagtttatc agggcattta cacaaaatac tgacctggta ccgtacgagg
     1441 ggttggaagt gcccgagggt ccccagcccg tggctaggcc acacattgaa gatgatgtca
     1501 ttatgcaggc tgttatgata tccctggggg cagacctgct accgctggcg gtgcaggctt
     1561 caaccgggga caattataac gtggccaggt actttgtgat accgggaaga tgcaccatgg
     1621 aacggtggcc ctggaactgt gccagacagg cattcgggat ccacggagca tacacccacg
     1681 tccacagcag cgtgcagagg ggtattcgcg gccttggcaa cctgttgttt cacagcaccc
     1741 tgtttccagg cggacagacc cagggggccc tcaccggcct gtatgccacc gaaccggccc
     1801 tgggaccccg tgcgcacagc cgatttcgtc gcatattcgc caagggcgta cagcaggccg
     1861 agatgctgca gggagcggga gtccccaccc tggggggttt cttaaaaacg gtgcgcacca
     1921 tcgccaccac tcctggcaac gccctggcag tctgctccat ctctaccact acttccaaag
     1981 aatgcatctc cctgagaagg atgatccccc agcagacagt ggtgtgtctg ggcaggtttg
     2041 agcccacgga cggaccggac acctacccta acctctatcg cgacagctcc gacaacgcgg
     2101 tgcgcatctt ggagaccctg aagctggttc agcggcttgc caagggccct atcttctccg
     2161 gactaaacag atcgcatgac ccggccccgg tggtgaggca cctgcaggcg ctagcgccgc
     2221 ggactggcct ggagctgttt gtctccaagc tacccgacga ggtgcgcgcc cacctgccag
     2281 cggatcccgc tgccggtccg gatgccgtgg aggcggcggt ggcagagcac tttctcaacg
     2341 tgtattgctc cctggtgttt gcggtggtgg cagagtcggg cgcggtgcct ggggatctgg
     2401 gcgagacccc gttggaggta ctgcagcgcg ccgcgcgcct gtgcgcgtgc cagataacgg
     2461 tcctcgggag gacctcggag cagccaggca tcagaatagt agacgacctg accggggaga
     2521 ccacgagggt cttctctgta gaccagccct cgtccacccc cccgtccccc tggctggcgc
     2581 tgtccgatgg tgtccgcgtc tcggggcacc ccgaggatgt tgactggggc ctttttgcca
     2641 ccggctccac aatccaccag ttacttcgcc acgcgacggt tggcagcaag gagttcttta
     2701 cgcgacacat ggacagatgc tccaacggcc tcatcgccca acaggctggc gtgggacccc
     2761 tggacatacc ggtttcagac taccacctgg tgctgcactc gtccatgctg gccgagaggg
     2821 tggcgcccag agtgcccgac acggtggagg ccatcactcc gtccatggcc aacctcctac
     2881 acaaggactt cgagacctgg gtgaaggccc tgccccagga gctgcttcca gtgccagcgt
     2941 ggaggggtca ggccatggcc atgggggagc aggcctacaa gatggctact aatgtatcca
     3001 ccggggccac ctatgccatc accgaggctc tcaccaacct catgttcagt cccgtgtcca
     3061 agctccagga cgtagtgcta actggcgccg tggcgtggag tccggaggac caccaagccg
     3121 gcctcctaca ggagtgcctc ttcgcctgca aggaattctg ccgggagctg ggagtggcac
     3181 tgtccatctc ctcggctgcc agctctccga cgctttcgga gcgccatgtg cgcatcacac
     3241 aacagcaaga aacggtggag gtccttccct tcaactcggt ggtgtttacc agctgggccg
     3301 aggtcaaggg atccagatac agggtcaccc cggacgtaaa ggtcgaaggc aacgccctgg
     3361 tatacctggc cgtgaatcag agctgtctca tagccgggtc caccttcgag cacaacttcc
     3421 tggcatccag gcacccaata ccccctctga acccgtccac ggtcgccagc ctgttcatgc
     3481 ttgttaagta cctgatgtcc aagaggctca ttgtatcggg gcacgacata ggggacgggg
     3541 ggcttctccc atctgcaatc gagatggccc tggccggctg caggggactg cagctctcac
     3601 tacccgccca ccctaacccg ctcgaactta tggtttcaga gacccctggg gcactggttg
     3661 aggtgcccca ggtacacttg tcagaggtgc tgcgggcggc tagggactac cgctgcgtgg
     3721 cacacccact gggcaccgtt ggccccgagg gacaaggcaa caacgtcacg gttttgcaga
     3781 acgagacagt tgtgtttcaa gagaccctga cttccttgca agtctcatgg acctcctttt
     3841 ctgacgaaat gtggaacctg gtgacgcctc ccctgcaccc actggaggac atgcacagga
     3901 aggacctggg acgtctggag catcacctgg gcagcctaag ggccatgtgc cttgggagtc
     3961 agctgcgcct gttttcatgc cccacctccc cgcgccgtgt ggccgcgttg gtgctgcctg
     4021 ggagcagcgc cccgtacgcg ctcatggccg cgttgcagaa cacgggcttt gaggtggcca
     4081 cggtgactgt ggaagagctt aaacgaggac agtccttgtc ggggttctct ggtctgatta
     4141 catgtctcag aacaggctgc caggccagct acgccagcgc caggggatgg gtcctggcgc
     4201 tgtgtaacga ccctacctgt gcctccaccc tgacagagtt tctaaacagg cccgacacat
     4261 tctccatctg ctgcggcgaa gtgggcttcc agcttctggt ggccctgggt gtagtgggcc
     4321 ggtcggaatc ctcaccatac acgtacggac ccacaccacc ccagcgctgg gcggtaaacc
     4381 tggagaccaa cgtgtccaag ctgtatgaca gccactggct aaacatacag atccctcaga
     4441 acactaagag cgttttcctc cgagtgttgc gggggacggt gctgcccagc tgggcccagg
     4501 gcgagtacct gggggtccgg tacgagcagg acgccctcga gtacatactg agacagcgag
     4561 gcgagataac cctcacctac catggaaatg ccgcggatga gaccctgcca gccagacact
     4621 atcccagaaa ccccacaggc aactccacgg tggccggact tacatccagt gacgggcgac
     4681 acgctgccct gatcatagac ccatccctga tgttccatcc gtggcagtgg cagcatgttc
     4741 caccagacct aacacccctg tccatgtccc cgtgggccat ggcgttccag tcaatctacc
     4801 tatggagcgt caagaagatc aacgaccacc actcagggcg gccgctcgag ggaggcggtg
     4861 gatggagcca cccccagttc gagaagggcg gcggcagcgg cggcggcagc ggcggcggca
     4921 gctggagcca cccccagttc gagaaggggc ccgtttaaac ccgctgatca gcctcgactg
     4981 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     5041 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     5101 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     5161 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     5221 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     5281 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5341 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5401 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5461 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5521 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5581 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5641 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     5701 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5761 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5821 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5881 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5941 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     6001 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     6061 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     6121 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     6181 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     6241 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     6301 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     6361 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     6421 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     6481 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     6541 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6601 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     6661 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6721 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6781 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8941 ccccgaaaag tgccacctga cgtc
//
