LOCUS       sequence-267582-        7049 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF50.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  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 7049)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ORF50-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..7049
                     /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             2948..2971
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3008..3031
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3057..3074)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3063..3287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3333..3761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3420..3439)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3630..3651
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3770..3790)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3775..4104
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3955..4090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4017..4036
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4152..4199
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4218..4592
                     /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    4722..4843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4759..4778)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4813..4832
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4892..4908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4892..4908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4905..4927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4916..4932
                     /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(4940..4946,4947..4964,4965..4970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4985..5006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5123..5140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5294..5882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5374..5393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6053..6844,6845..6913))
                     /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     6676..6695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6914..7018)
                     /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 attcatgaaa gaatgttcca agcttggtgc gctatgtggt ctgtttacag acattaacat
     1081 gtttaacctt ttctgctttt ttcgtgcctc tcgaatgagg accaaaggcg cggccgggta
     1141 caacgtgcca tgcgcagagg catcccaagg cattattcgg atcctcacgg agaggatctt
     1201 attctgcaca gaaaaggcat ttctgacagc cgcatgcagc ggggtgagcc tgcctccagc
     1261 catatgtaag ctactacacg aaatatacac tgaaatgaag gccaaatgcc tgggggcctg
     1321 gaggcgactc gtctgcaatc ggaggcccat tatgatatta acctcttccc tactgaagct
     1381 ctacaacacg tacgataccg ccgggctgct ctctgagcag tccagggccc tctgcctttt
     1441 ggttttccaa ccggtctacc ttccgaggat tatggcgccg ctggagatca tgaccaaggg
     1501 tcagctcgcc cctgaaaact tttacagcat caccggttct gctgagaaac gccggccaat
     1561 taccaccggc aaggtcactg gactgtccta tccaggaagc ggtctcatgc cagaatcttt
     1621 aattttgcca atcctggagc caggactgtt gccggcttcc atggtagacc tcagcgatgt
     1681 gctggcaaaa cccgccgtta ttctgagcgc ccctgccctg agccagtttg tcattagcaa
     1741 accccatccc aacatgccgc acaccgtcag catcatcccc tttaacccat cgggtacaga
     1801 cccggcgttt attagtacgt ggcaggccgc gtcacagaat atggtgtaca acacatccac
     1861 cgcgccctta aaaccggcca ccggtagttc acagacggtg tcagtcaagg cggttgctca
     1921 aggggccgtg attactgcga caacggtgcc gcaggcaatg ccagcgcggg gtaccgaagg
     1981 ggagttgcct gtaatgtcag cgtccactcc tgcaagagat caggtcgctg catgttttgt
     2041 cgcagagaac accggagatt ctcccgacaa cccgagctct ttcctgacgt catgtcaccc
     2101 ttgcgatccg aacacggtta tagtggccca gcaatttcaa ccaccgcaat gcgttacgtt
     2161 gttgcaggtt acctgtgccc cctcttcgac accacccccc gattcaacag tccgggcccc
     2221 ggtggtgcag ttgccaacag tagtccctct gccggccagc gcgttcctcc cggcgctcgc
     2281 ccaaccagaa gcctcgggcg aagagcttcc gggcggtcat gacggagacc aaggtgtgcc
     2341 gtgtagagat tcaacggcgg cggctacggc ggcagaggcg acaacaccca aacgaaagca
     2401 gagaagcaaa gagaggagct caaagaagcg taaggctttg accgtgccag aagccgacac
     2461 cacgccatcg accacgacac ctggtacctc tttgggatca attaccaccc cccaggatgt
     2521 gcacgccacg gatgtcgcca cgtctgaggg accatcggag gcacaacccc cgctactgtc
     2581 gttacccccg ccactggacg tagatcagag tctattcgcc ctgttagacg aagcgggccc
     2641 tgaaacatgg gatgtcgggt cgcctctctc ccccactgac gacgcgctgt tgtccagtat
     2701 tctgcaagga ctgtaccagc tggacacgcc accgcctctg cggtcaccct cccccgcttc
     2761 cttcggcccg gagtctccgg cggatatacc gtcaccttct ggtggagagt atacgcaact
     2821 gcaaccggtc agggcgacct cggcgacgcc cgctaacgag gtacaggagt ccggcacact
     2881 gtaccagctg caccaatggc gtaattactt ccgagacgga gggcggccgc tcgagggagg
     2941 cggtggatgg agccaccccc agttcgagaa gggcggcggc agcggcggcg gcagcggcgg
     3001 cggcagctgg agccaccccc agttcgagaa ggggcccgtt taaacccgct gatcagcctc
     3061 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3121 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3181 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3241 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3301 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3481 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3661 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3721 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3781 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3841 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4021 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4081 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4141 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     4201 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     4261 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     4321 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     4381 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     4441 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     4501 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     4561 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     4621 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4681 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4741 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4801 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4861 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5821 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5881 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5941 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6001 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6061 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6121 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6181 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6241 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6301 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6361 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6421 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6481 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6541 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6601 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6661 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6721 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6781 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6841 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6901 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6961 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7021 catttccccg aaaagtgcca cctgacgtc
//
