LOCUS       sequence-267609-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF59.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  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 6344)
  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-ORF59-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..6344
                     /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             2243..2266
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2303..2326
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2352..2369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2358..2582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2628..3056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2715..2734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2925..2946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3065..3085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3070..3399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3250..3385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3312..3331
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3447..3494
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3513..3887
                     /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    4017..4138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4054..4073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4108..4127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4187..4203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4187..4203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4200..4222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4211..4227
                     /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(4235..4241,4242..4259,4260..4265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4280..4301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4418..4435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4589..5177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4669..4688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5348..6139,6140..6208))
                     /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     5971..5990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6209..6313)
                     /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 attcatgcct gtggattttc actatggggt cagggtggac gtgaccctcc tgtctaaaat
     1081 caggagggtc aatgaacaca ttaagagtgc cactaaaacc ggagtagtgc aagtgcacgg
     1141 atcggcttgc acgccaaccc tcagtgtgct gtccagcgtg gggacagctg gcgttctggg
     1201 gttaagaata aagaatgccc ttacgcccct ggtgggacac acggaaggca gtggagacgt
     1261 tagcttcagc ttcaggaata cgtccgtcgg tagcggcttc acgcacacgc gtgagctatt
     1321 cggtgcgaat gtactcgacg ctggcatagc cttttatcga aagggggagg catgcgatac
     1381 gggggcccaa ccgcagttcg tcaggaccac catttcctac ggggacaacc tcacctccac
     1441 cgtgcacaaa agcgtggtgg accaaaaggg tatcctacct ttccacgatc ggatggaagc
     1501 cggtggtagg accaccaggc ttctcctctg tggcaaaacg ggtgcattcc tacttaagtg
     1561 gctaaggcag caaaaaacaa aagaggacca aacggtgact gtgtctgtca gcgagacgct
     1621 gtcgatcgtg acattttctc ttggtggcgt gagcaagatc attgatttca agccagaaac
     1681 caaacccgtt tctggctggg atggtctgaa ggggaagaag tcggtggatg tgggcgtggt
     1741 gcataccgac gccctgagcc gggttagcct ggagtcctta atcgccgccc tgagactgtg
     1801 taaagtcccg ggttggttta cccccgggct gatttggcac tccaacgaaa tattagaagt
     1861 ggaaggtgtg cccactggct gccaatcagg tgacgtaaag ttaagtgtcc tacttttaga
     1921 ggtaaaccga tctgtgtctg ccgagggagg cgagtcttcg caaaaggttc ccgattctat
     1981 accggactcc aggaggcagc cggaattgga gtctccggat tctccccctc tcacaccagt
     2041 tggtcccttt gggccgctgg aggacgcctc tgaggacgct gcctctgtca ccagctgccc
     2101 cccagcggcg ccaactaagg acagcacaaa gaggcctcac aagaggcgct cagactcgag
     2161 ccagtccagg gatcgtggga aggtgcccaa aaccacattt aaccccctga ttggagggcg
     2221 gccgctcgag ggaggcggtg gatggagcca cccccagttc gagaagggcg gcggcagcgg
     2281 cggcggcagc ggcggcggca gctggagcca cccccagttc gagaaggggc ccgtttaaac
     2341 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2401 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2461 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2521 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2581 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2641 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2701 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2761 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2821 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2941 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3001 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3301 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3421 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     3481 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     3541 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     3601 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     3661 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     3721 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     3781 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     3841 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     3901 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3961 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4141 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4201 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4261 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4321 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4381 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4441 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4501 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4561 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4621 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4681 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4741 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4801 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4861 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4921 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4981 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5041 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5101 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5161 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5221 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5281 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5341 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5401 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5461 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5521 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5581 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5641 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5701 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5761 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5821 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5881 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5941 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6001 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6061 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6121 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6181 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6241 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6301 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
