LOCUS       sequence-291853-        5924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) K5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  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 5924)
  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-K5-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..5924
                     /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             1823..1846
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1883..1906
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1932..1949)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1938..2162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2208..2636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2295..2314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2505..2526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2645..2665)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2650..2979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2830..2965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2892..2911
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3027..3074
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3093..3467
                     /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    3597..3718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3634..3653)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3688..3707
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3767..3783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3767..3783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3780..3802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3791..3807
                     /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(3815..3821,3822..3839,3840..3845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3860..3881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3998..4015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4169..4757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4249..4268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4928..5719,5720..5788))
                     /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     5551..5570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5789..5893)
                     /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 attcatggcg tctaaggacg tagaagaggg tgtagaggga cccatctgct ggatatgtag
     1081 ggaagaggtg gggaacgagg gcatacaccc ctgcgcctgt accggagagc tggatgtcgt
     1141 ccacccgcag tgtttaagca cttggctaac agtgtctcga aacacggcct gtcaaatgtg
     1201 tcgcgttata taccgcacgc gcacgcagtg gcgtagtcgc cttaacctgt ggccggagat
     1261 ggagcgccaa gaaatttttg aactgtttct gctgatgtct gtggtggtgg ctgggcttgt
     1321 cggcgtggcg ttgtgcacct ggacgctcct ggtcatccta actgctcctg cgggaacatt
     1381 ctcccccggg gccgtgctcg gttttctctg cttctttggg ttttaccaaa tatttattgt
     1441 gtttgcattt ggcggcatat gccgcgtaag tggcactgtg agggcattat acgcggcaaa
     1501 taacacccgg gtgaccgtac tgccataccg acggccgcgc cggccaaccg cgaacgaaga
     1561 taacatcgaa ttgacggtcc ttgtcggacc cgcgggcggg acggacgagg agcccacgga
     1621 cgagtcatct gaaggagacg tcgcctctgg agacaaagaa cgtgacggtt catccggaga
     1681 cgagccggac gggggcccga acgaccgtgc gggacttagg gggacagcgc gtaccgacct
     1741 atgcgcgccc acaaaaaagc cggtgcggaa aaatcatcca aaaaacaacg gtggagggcg
     1801 gccgctcgag ggaggcggtg gatggagcca cccccagttc gagaagggcg gcggcagcgg
     1861 cggcggcagc ggcggcggca gctggagcca cccccagttc gagaaggggc ccgtttaaac
     1921 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1981 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2041 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2101 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2161 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2221 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2281 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2341 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2401 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2461 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2521 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2581 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2641 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2701 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2761 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2821 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2881 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2941 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3001 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     3061 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     3121 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     3181 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     3241 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     3301 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     3361 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     3421 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     3481 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3541 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3601 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3661 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3721 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3781 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3841 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3901 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3961 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4021 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4081 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4141 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4201 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4261 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4321 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4381 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4441 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4501 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4561 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4621 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4681 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4741 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4801 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4861 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4921 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4981 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5041 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5101 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5161 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5221 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5281 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5341 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5401 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5461 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5521 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5581 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5641 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5701 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5761 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5821 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5881 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
