LOCUS       sequence-267436-        7323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF22.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7323)
  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 7323)
  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-ORF22-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..7323
                     /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             3222..3245
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3282..3305
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3331..3348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3337..3561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3607..4035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3694..3713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3904..3925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4044..4064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4049..4378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4229..4364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4291..4310
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4426..4473
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4492..4866
                     /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    4996..5117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5033..5052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5087..5106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5166..5182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5166..5182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5179..5201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5190..5206
                     /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(5214..5220,5221..5238,5239..5244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5259..5280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5397..5414)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5568..6156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5648..5667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6327..7118,7119..7187))
                     /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     6950..6969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7188..7292)
                     /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 catgcagggt ctagccttct
     1021 tggcggccct tgcatgctgg cgatgcatat cgttgacatg tggagccact ggcgcgttgc
     1081 cgacaacggc gacgacaata acccgctccg ccacgcagct catcaatggg agaaccaacc
     1141 tctccataga actggaattc aacggcacta gtttttttct aaattggcaa aatctgttga
     1201 atgtgatcac ggagccggcc ctgacagagt tgtggacctc cgccgaagtc gccgaggacc
     1261 tcagggtaac tctgaaaaag aggcaaagtc tttttttccc caacaagaca gttgtgatct
     1321 ctggagacgg ccatcgctat acgtgcgagg tgccgacgtc gtcgcaaact tataacatca
     1381 ccaagggctt taactatagc gctctgcccg ggcaccttgg cggatttggg atcaacgcgc
     1441 gtctggtact gggtgatatc ttcgcatcaa aatggtcgct attcgcgagg gacaccccag
     1501 agtatcgggt gttttaccca atgaatgtca cggccgtcaa gttttccata tccattggca
     1561 acaacgagtc cggcgtagcg ctctatggag tggtgtcgga agatttcgtg gtcgtcacgc
     1621 tccacaacag gtccaaagag gctaacgaga cggcgtccca tcttctgttc ggtctcccgg
     1681 attcactgcc atctctgaag ggccatgcca cctatgatga actcacgttc gcccgaaacg
     1741 caaaatatgc gctagtggcg atcctgccta aagattctta ccagacactc cttacagaga
     1801 attacactcg catatttctg aacatgacgg agtcgacgcc cctcgagttc acgcggacga
     1861 tccagaccag gatcgtatca atcgaggcca ggcgcgcctg cgcagctcaa gaggcggcgc
     1921 cggacatatt cttggtgttg tttcagatgt tggtggcaca ctttcttgtt gcgcagggca
     1981 ttgccgagca ccgatttgtg gaggtggact gcgtgtgtcg gcagtatgcg gaactgtatt
     2041 ttctccgccg catctcgcgt ctgtgcatgc ccacgttcac cactgtcggg tataaccaca
     2101 ccacccttgg cgctgcggcc gccacacaaa tagctcgcgt gtccgccacg aagttggcca
     2161 gtttgccccg ctcttcccag gaaacagtgc tggccatggt ccagcttggc gcccgtgatg
     2221 gcgccgtccc ttcctccatt ctggagggca ttgctatggt cgtcgaacat atgtataccg
     2281 cctacactta tgtgtacaca ctcggcgata ctgaaagaaa attaatgttg gacatacaca
     2341 cggtcctcac cgacagctgc ccgcccaaag actccggagt atcagaaaag ctactgagag
     2401 catatttgat gctcacatca atgtgtacca acatagggct gggcgaaatg atcgcccgct
     2461 tttccaaacc ggacagcctt aacatctata gggcattctc cccctgcttt ctaggactaa
     2521 ggtacgattt gcatccagcc aagttgcgcg ccgaggcgcc gcagtcgtcc gctctgacgc
     2581 ggactgccgt tgccagagga acatcgggat tcgcagaatt gctccacgcg ctgcacctcg
     2641 atagcttaaa tttaattccg gcgattaact gttcaaagat tacagccgac aagataacag
     2701 ctacggtacc cttgcctcac gtcacgtata tcatcagttc cgaagcactc tcgaacgctg
     2761 ttgtctacga ggtgtcggag atcttcctca agagtgccat gtttatatct gctatcaaac
     2821 ccgattgctc cggctttaac ttttctcaga ttgataggca cattcccata gtctacaaca
     2881 tcagcacacc aagaagaggt tgcccccttt gtgactctgt aatcatgagc tacgatgaga
     2941 gcgatggcct gcagtctctc atgtatgtca ctaatgaaag ggtgcagacc aacctctttt
     3001 tagataagtc acctttcttt gataataaca acctacacat tcattatttg tggctgaggg
     3061 acaacgggac cgtagtggag ataaggggca tgtatagaag acgcgcagcc agtgctttgt
     3121 ttctaattct ctcttttatt gggttctcgg gggttatcta ctttctttac agactgtttt
     3181 ccatccttta ttcggggcgg ccgctcgagg gaggcggtgg atggagccac ccccagttcg
     3241 agaagggcgg cggcagcggc ggcggcagcg gcggcggcag ctggagccac ccccagttcg
     3301 agaaggggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3361 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3421 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3481 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3541 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3601 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3661 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3721 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3781 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3841 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3901 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3961 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4021 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4141 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4201 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4261 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4321 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4381 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
     4441 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
     4501 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     4561 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     4621 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     4681 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     4741 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     4801 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     4861 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4921 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4981 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5041 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5101 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5161 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5221 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5281 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5341 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5401 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5461 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5521 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5581 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5641 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5701 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5761 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5821 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5881 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5941 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6001 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6061 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6121 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6181 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6241 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6301 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6361 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6421 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6481 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6541 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6601 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6661 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6721 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6781 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6841 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6901 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6961 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7021 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7081 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7141 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7201 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7261 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7321 gtc
//
