LOCUS       sequence-267620-        7917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF63.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7917)
  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 7917)
  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-ORF63-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..7917
                     /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             3816..3839
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3876..3899
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3925..3942)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3931..4155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4201..4629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4288..4307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4498..4519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4638..4658)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4643..4972
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4823..4958
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4885..4904
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5020..5067
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5086..5460
                     /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    5590..5711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5627..5646)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5681..5700
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5760..5776)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5760..5776)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5773..5795)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5784..5800
                     /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(5808..5814,5815..5832,5833..5838))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5853..5874
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5991..6008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6162..6750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6242..6261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6921..7712,7713..7781))
                     /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     7544..7563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7782..7886)
                     /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 catggacggc acagacgctc
     1021 ttgaaaagct taccaaaggc ctgagtgggg gaggcgggag ccttcaccag acaaagctgt
     1081 tgatggaatt tcaactccga ggactgccgg tgcctgccct cttaaacagc agcacaacag
     1141 agcagttttt aaatactgtt gcccaactgc cgacggacct atcaaaattt atacgcgact
     1201 atcgcgtgtt cgcactggtt cgcgcggcgt attttttaga acccccttct agcatcgacc
     1261 cccttgaggc agcgcgcgct cttggacgcc tggttgatat attatcatca aaaccaccgc
     1321 agaacaccgc accggcgcag ccacccacct ccgacgacac cctgaataac tgtacattgc
     1381 tcaaactact agcccactac gcggatcaga tagcaggttt caaaaccccc gctctccctc
     1441 ccgtgccacc tggaatcatc ggcctgttca catgcgtgga acagatgtac cacgcatgtt
     1501 ttcagaaata ctgggcagct gcactacccc caatgtggat actgacatac gaccctccca
     1561 cttctccgtt acaggactgg cttatagtcg cctatggtaa caaggaagga ctgctactcc
     1621 cctctggcat accctcggag gaggtgttag ccaaaacatt agtaacagaa caccacgagt
     1681 tgttcgtatc gcggtcgaat tcgaccgaga ccgccgtcac catgcccgta tccaaagaac
     1741 gcgccctcgc catctaccgg gtgttcgcca agggtgaggt ggtggcggaa aatactccca
     1801 ttcttgcctt caccgacgtg gaactatcca cactcaaacc ccactatctg ttcatctatg
     1861 attttatcat agaggcatta tgcaagagct acacatactc atgcacccag gcccgcctgg
     1921 aatccttttt gagccgaggt atagacttca tgactgacct aggtcagtac ctagataccg
     1981 ctactagcgg caagcagcag ctgacgcaca gccaaataaa ggaaatcaaa tacaggctgc
     2041 taagctgcgg tctctcggct tccgcgtgtg atgttttcag aactgtgatc atgaccctcc
     2101 catatcgacc gacccccaac ctcgctaacc tgtccacgtt tatggggatg gttcaccaac
     2161 tgaccatgtt cggacactat ttctaccggt gcctgggcag ctacagtccc accggcttgg
     2221 ccttcacaga attgcaaaag atactgacac gcgccagcgc ggagcaaacg gaacgtaacc
     2281 cgtggagaca tccgggtatc tcggacattc cactgcgttg gaaaatatcg cgtgctctag
     2341 cattcttcgt ccctccggcc cccataaaca ctttgcagcg cgtgtacgcc gcgctgccct
     2401 cgcaactcat gcgggccatc ttcgagatct cggtcaagac cacatgggga ggcgccgtac
     2461 cggcaaacct ggcgcgcgac attgacacag gaccgaacac acaacatatc tcctccacac
     2521 caccgcccac cctcaaggat gttgagacat actgtcaagg tctgcgggtg ggagacacgg
     2581 agtacgatga ggacattgtg agaagcccgc tctttgcaga cgcgtttacc aagagtcact
     2641 tgttgcctat actgcgcgag gttctggaaa accgcctgca gaaaaacaga gctctgtttc
     2701 agataagatg gctgataata tttgctgccg aggcggcaac cgggctcatc cctgccaggc
     2761 gcccgctagc cagagcctac ttccacatca tggacattct ggaggagaga cattcccaag
     2821 acgccctata caaccttttg gactgtatcc aggagctctt cacccacatc aggcaggctg
     2881 ttccagacgc acagtgtccg cacgcctttc tacagtccct gttcgtcttt caattccgcc
     2941 ctttcgtact caaacaccag cagggtgtaa ccttgtttct agatggcttg cagacatccc
     3001 tccccccggt gataagtctg gccaaccttg gagacaagct gtgtcgtctc gagttcgagt
     3061 acgacagcga gggcgacttc gtgcgcgtgc cagttgcacc gccagaacaa ccaccgcacg
     3121 tacatctgtc gcatttcaag aagacaatac agaccatcga acaggccacc agggaggcca
     3181 ccgtagccat gacaacaatc gcaaagccaa tataccccgc ctacatccgg ttactgcagc
     3241 ggctagaata tcttaacaga ctcaaccacc acattctcag gattcccttc ccacaggacg
     3301 ccctttctga actccaggaa acctacctgg cggcgtttgc acggttgaca aaattggcag
     3361 cggacgcagc aaacacttgt agctactccc tcaccaagta ctttggagtt ttattccaac
     3421 accagctggt ccccacggcc atcgttaaaa aactgctaca tttcgacgag gctaaagata
     3481 ccacagaagc ctttttacag agcctggcac aacccgtagt gcagggacaa cggcaggggg
     3541 cggctggcgg gtcgggtgtc ctgacgcaga aagaacttga gctcttgaac aaaataaacc
     3601 cacagtttac agacgctcag gctaacattc ctccatctat taaacgttca tattcaaata
     3661 aatatgacgt ccctgaggtc tcagtcgact gggaaacgta ctcccggtct gccttcgagg
     3721 caccggacga cgaactccgt tttgtcccac tgacgctggc aggcctccgg aaactgtttg
     3781 tcgaatcggg gcggccgctc gagggaggcg gtggatggag ccacccccag ttcgagaagg
     3841 gcggcggcag cggcggcggc agcggcggcg gcagctggag ccacccccag ttcgagaagg
     3901 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3961 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4021 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4081 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4141 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     4201 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4261 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4321 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4381 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4441 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4501 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4561 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4621 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4681 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4741 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4801 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4861 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4921 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4981 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     5041 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     5101 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     5161 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     5221 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     5281 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     5341 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     5401 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     5461 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5521 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5581 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5641 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5701 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5761 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5821 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5881 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5941 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6001 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6061 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6601 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6841 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6901 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6961 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7021 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7081 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7141 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7201 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7261 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7321 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7381 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7441 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7501 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7561 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7621 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7681 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7741 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7801 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7861 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
