LOCUS       sequence-291860-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) K8a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  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 5867)
  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-K8a-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..5867
                     /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             1766..1789
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1826..1849
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1875..1892)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1881..2105
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2151..2579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2238..2257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2448..2469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2588..2608)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2593..2922
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2773..2908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2835..2854
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2970..3017
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3036..3410
                     /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    3540..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3577..3596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3631..3650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3710..3726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3710..3726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3723..3745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3734..3750
                     /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(3758..3764,3765..3782,3783..3788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3803..3824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3941..3958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4112..4700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4192..4211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4871..5662,5663..5731))
                     /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     5494..5513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5732..5836)
                     /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 attcatgccc agaatgaagg acatacctac taagagttcc ccgggaacgg acaattctga
     1081 gaaagatgaa gctgtcattg aggaagatct aagcctcaac gggcaaccat tttttacgga
     1141 caatactgac ggtggggaaa acgaagtctc ttggacaagc tcgctgttgt caacctacgt
     1201 aggttgccag cccccggcca taccggtctg tgaaacggtc attgacctta cagcgccttc
     1261 ccaaagtggc gcgcccggtg acgaacatct gccatgctca ctgaatgcag aaactaaatt
     1321 ccacatcccc gatccttcct ggacgctctc tcacacacca ccaagaggac cacacatttc
     1381 gcaacagctt ccaactcgca gatccaagag gcgactacat agaaagtttg aagaggaacg
     1441 cttatgcacc aaggccaaac agggcgcagg tcgccccgtg cctgcgtctg tagttaaggc
     1501 cgaagtatgt gatcagtcac attctcccac gcgaaagcaa ggcagatacg gccgcgtgtc
     1561 atcgaaagca tacacaagac agctgcagca ggcattagaa gaaaaggatg cacaactatg
     1621 tttcctagcg gcgagattgg aggcacataa ggaacagatt attttccttc gcgacatgct
     1681 gatgcgaatg tgccagcagc cagcgtcgcc aacggacgcg ccactcccac catgttcagg
     1741 gcggccgctc gagggaggcg gtggatggag ccacccccag ttcgagaagg gcggcggcag
     1801 cggcggcggc agcggcggcg gcagctggag ccacccccag ttcgagaagg ggcccgttta
     1861 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1921 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1981 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2041 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2101 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2161 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2221 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2281 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2341 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2401 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2461 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2521 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2581 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2641 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2821 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2881 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2941 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     3001 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     3061 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     3121 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     3181 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     3241 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     3301 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     3361 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     3421 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3481 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3661 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3721 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3781 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3841 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3901 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
