LOCUS       sequence-267598-        7685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF56.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7685)
  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 7685)
  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-ORF56-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..7685
                     /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"
     regulatory      2799..2808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3584..3607
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3644..3667
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3693..3710)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3699..3923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3969..4397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4056..4075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4266..4287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4406..4426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4411..4740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4591..4726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4653..4672
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4788..4835
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4854..5228
                     /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    5358..5479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5395..5414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5449..5468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5528..5544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5528..5544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5541..5563)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5552..5568
                     /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(5576..5582,5583..5600,5601..5606))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5621..5642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5759..5776)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5930..6518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6010..6029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6689..7480,7481..7549))
                     /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     7312..7331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7550..7654)
                     /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 attcatggag acgacatacc gccgcgaggc gctgacagta agggttattt tttgtacgag
     1081 tggcgacagc gccgagacga tcgccgacgt ccttacgggg gccccaacgt cagcgtcctt
     1141 cttttctgta ctccacgacc ttttttattc ccagatactc gcccccaggg taaccctaaa
     1201 attgtgcctc cccgcacggc gtcctggcaa cggcacaagg tgttcgcccg tgttggtcct
     1261 acgtactgac gcatcagtgg cctcggggtt ccttggcggc cggccactgg aggcgtccga
     1321 cattaaatat atgctgctca gcgaccagac cgcggggttg ttcaagccgc tgttggagat
     1381 aatcggtggc gcgcgcgcac caccaaatca ggacgcgtgc actttccaga gccaggtggc
     1441 ctggctcaga acgaaatttg ttaccgcatt gagaaaactt tacaagatga ctccctcacc
     1501 ctactggatg ctgtctgcat ttggcgctca ggaagcccag ttcgtcctga ccagctcatt
     1561 ctattttttt gaacacactg tggtctgtac cacagagaca gtttctcacc tgtctagact
     1621 gttttcgcct caacagggac agacgctggt ttccgttacc agccacgagg agctggggca
     1681 gctatacggc acttcccctt tcaggcggcg cgtccccgcg ttcgtcgctt atgtaaaaga
     1741 gaaattagcg agagacagtc tggagacgga ggccatcgac cgcaccatag accagatcag
     1801 gggcaaactc atgctgtcta accaggacct ggtccatttc atatatatct ccttttatca
     1861 gtgcctcaac aaacgggcgt tcctgcgcta ctctagacag acgtcctctt caagtgctct
     1921 aagggagctg ggggaagacc ctcaattgtg tggcgcccta cacggggagt ttcgtgacca
     1981 cgtccagtcc tactaccaca aaaaaaccta cctatccact tacatagaca ttcggtacgt
     2041 gggtggcgta ttaccagacg gctattttgg cgggagtctt gtaggcgagc ggtgcgttta
     2101 ttggtgcggg cagtcaaagg acacggccag cctgttggcc accattagcc aacaggtgcc
     2161 gcacctgagg ttgcaaaacg agttcgctgg catgctagac gtggccgcac tgcgaggttc
     2221 cgatgacggt cagtttaaag agggcctttt ctcccacagt caagccctac ccctgtacag
     2281 gtgcgagttt ctgggcaagc agtttttcac aatgcttcag gaagacggcc tagagcgata
     2341 ctgggagcaa agtgtgatat ttccaggcga ccaggactgg gatatgttat ctgacaaaga
     2401 cctcacctac cgaatttttt accatgacct cagcctatcg ctgccaacac tgaaggaaca
     2461 gctccttgtt tcaagacacg aatacttcaa ccctcgcttg ccagtgtata gatgggtatt
     2521 agactttgac ctgcccgtct gccgcgacat tgacaggaca ttcgaggagg tgcactctct
     2581 ctgttgttcc ctgcgtgagg ccatactcga catcattcaa ctccttggac cagtggatcc
     2641 tcgaacacac ccagtatatt ttttcaaatc agcctgtcca ccggacgagt ggcgcggcga
     2701 agacgtcgcc agcaccagct tctgtcggtg tcatgacaaa ctgggtatgc gtattatcgt
     2761 cccgttccca gaaggagtat gcgtcgttgg gtcggagccc atggtggcac tcactggcat
     2821 tctaaacagg acgataaagc ttgatccgga gctggtccac agattcccgt caatacaaaa
     2881 aaaggggggc cctttcgact gtggcatata cggccgagga cgaagcgtcc ggcttcccca
     2941 ctgttacaag gtgggcttag tgggggaact ctgccgccta ctgaagatac tagtctgtca
     3001 ccccgccccc aacggcaagg cgcagtacgt gcggcgcgcc tttacgcttc gcgaactgct
     3061 ccatcactcc ccgggccaca gcgccggtca tgtcggccga atcatctata gcatcatgga
     3121 tcgcaatgag aattttttag aaaacaagac cattagctat ctgccggcca aaatacctca
     3181 catctttcag cggatagaga ccctatccgg tcgttcaata gaggactggc tacactcggc
     3241 cgtttgggat aaagcatacg acactatatg taaatttttc ccagatgaaa aagcacaaca
     3301 gttttctcac gttgcattta cgcaacaagg ggaaaacatc atccagttaa gaccccgtca
     3361 gggaagacac ttcctctgca tcaaccataa tcataaaaac aagtcaaaaa cagtccgtgt
     3421 attccttacc cttcattcca ttagggtgag cgaagtcacg gtaacactta tgagtcagtg
     3481 ttttgccagc aagtgtaaca ataatgttcc cacggcccat ttttcgtttg tggtaccagt
     3541 gggactggcc agttcagggc ggccgctcga gggaggcggt ggatggagcc acccccagtt
     3601 cgagaagggc ggcggcagcg gcggcggcag cggcggcggc agctggagcc acccccagtt
     3661 cgagaagggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3721 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3781 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3841 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3901 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3961 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4021 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4081 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4141 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4201 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4261 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4321 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4381 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4561 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4621 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4681 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4741 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     4801 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     4861 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     4921 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     4981 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     5041 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     5101 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     5161 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     5221 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5281 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5341 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5401 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5461 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5521 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5581 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5641 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5701 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5761 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5821 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5881 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5941 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6001 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6061 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6121 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6181 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6241 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6301 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6361 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6421 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6481 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6541 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6601 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6661 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6721 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6781 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6841 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6901 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6961 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7021 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7081 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7141 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7201 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7261 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7321 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7381 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7441 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7501 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7561 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7621 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7681 acgtc
//
