LOCUS       sequence-267434-        6896 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF21.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6896)
  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 6896)
  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-ORF21-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..6896
                     /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             2795..2818
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2855..2878
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2904..2921)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2910..3134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3180..3608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3267..3286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3477..3498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3617..3637)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3622..3951
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3802..3937
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3864..3883
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3999..4046
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4065..4439
                     /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    4569..4690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4606..4625)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4660..4679
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4739..4755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4739..4755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4752..4774)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4763..4779
                     /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(4787..4793,4794..4811,4812..4817))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4832..4853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4970..4987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5141..5729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5221..5240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5900..6691,6692..6760))
                     /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     6523..6542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6761..6865)
                     /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 attcatggca gaaggcggtt ttggagcgga ctcggtgggg cgcggcggag aaaaggcctc
     1081 tgtgactagg ggaggcaggt gggacttggg gagctcggac gacgaatcaa gcacctccac
     1141 aaccagcacg gatatggacg acctccctga ggagaggaaa ccactaacgg gaaagtctgt
     1201 aaaaacctcg tacatatacg acgtgcccac cgtcccgacc agcaagccgt ggcatttaat
     1261 gcacgacaac tccctctacg caacgcctag gtttccgccc agacctctca tacggcaccc
     1321 ttccgaaaaa ggcagcattt ttgccagtcg gttgtcagcg actgacgacg actcgggaga
     1381 ctacgcgcca atggatcgct tcgccttcca gagccccagg gtgtgtggtc gccctcccct
     1441 tccgcctcca aatcacccac ctccggcaac taggccggca gacgcgtcaa tgggggacgt
     1501 gggctgggcg gatctgcagg gactcaagag gaccccaaag ggatttttaa aaacatctac
     1561 caaggggggc agtctcaaag cccgtggacg cgatgtaggt gaccgtctca gggacggcgg
     1621 ctttgccttt agtcctaggg gcgtgaaatc tgccataggg caaaacatta aatcatggtt
     1681 ggggatcgga gaatcatcgg cgactgctgt ccccgtcacc acgcagctta tggtaccggt
     1741 gcacctcatt agaacgcctg tgaccgtgga ctacaggaat gtttatttgc tttacttaga
     1801 gggggtaatg ggtgtgggca aatcaacgct ggtcaacgcc gtgtgcggga tcttgcccca
     1861 ggagagagtg acaagttttc ccgagcccat ggtgtactgg acgagggcat ttacagattg
     1921 ttacaaggaa atttcccacc tgatgaagtc tggtaaggcg ggagacccgc tgacgtctgc
     1981 caaaatatac tcatgccaaa acaagttttc gctccccttc cggacgaacg ccaccgctat
     2041 cctgcgaatg atgcagccct ggaacgttgg gggtgggtct gggaggggca ctcactggtg
     2101 cgtctttgat aggcatctcc tctccccagc agtggtgttc cctctcatgc acctgaaaca
     2161 cggccgccta tcttttgatc acttctttca attactttcc atctttagag ccacagaagg
     2221 cgacgtggtc gccattctca ccctctccag cgccgagtcg ttgcggcggg tcagggcgag
     2281 gggaagaaag aacgacggga cggtggagca aaactacatc agagaattgg cgtgggctta
     2341 tcacgccgtg tactgttcat ggatcatgtt gcagtacatc actgtggagc agatggtaca
     2401 actatgcgta caaaccacaa atattccgga aatctgcttc cgcagcgtgc gcctggcaca
     2461 caaggaggaa actttgaaaa accttcacga gcagagcatg ctacctatga tcaccggtgt
     2521 actggatccc gtgagacatc atcccgtcgt gatcgagctt tgcttttgtt tcttcacaga
     2581 gctgagaaaa ttacaattta tcgtagccga cgcggataag ttccacgacg acgtatgcgg
     2641 cctgtggacc gaaatctaca ggcagatcct gtccaatccg gctattaaac ccagggccat
     2701 caactggcca gcattagaga gccagtctaa agcagttaat cacctagagg agacatgcag
     2761 ggtctcgggg cggccgctcg agggaggcgg tggatggagc cacccccagt tcgagaaggg
     2821 cggcggcagc ggcggcggca gcggcggcgg cagctggagc cacccccagt tcgagaaggg
     2881 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2941 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3001 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3061 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3121 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3181 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3241 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3301 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3361 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3421 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3481 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3541 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3601 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3661 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3721 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3781 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3841 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3901 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3961 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     4021 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
     4081 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     4141 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     4201 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     4261 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     4321 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     4381 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     4441 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4501 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4561 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4621 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4681 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4741 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4801 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4861 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4981 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5041 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5101 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5161 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5221 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5281 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5341 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5401 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5461 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5521 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5581 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5641 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5701 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5761 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5821 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5881 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5941 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6001 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6061 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6121 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6181 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6241 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6301 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6361 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6421 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6481 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6541 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6601 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6661 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6721 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6781 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6841 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
