LOCUS       sequence-436409-        8133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF73.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8133)
  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 8133)
  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-ORF73-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..8133
                     /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"
     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             4032..4055
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4092..4115
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4141..4158)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4147..4371
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4417..4845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4504..4523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4714..4735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4854..4874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4859..5188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5039..5174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5101..5120
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5236..5283
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5302..5676
                     /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    5806..5927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5843..5862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5897..5916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5976..5992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5976..5992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5989..6011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6000..6016
                     /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(6024..6030,6031..6048,6049..6054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6069..6090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6207..6224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6378..6966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6458..6477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7137..7928,7929..7997))
                     /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     7760..7779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7998..8102)
                     /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 tgccatggcg cccccgggaa
     1021 tgcgcctgag gtcgggacgg agcaccggcg cgcccttaac gagaggaagt tgtaggaaac
     1081 gaaacaggtc tccggaaaga tgtcaccttg gcgatgacct acatctacaa ccgcgaagga
     1141 agcatgtcgc cgactccgtc gacggccggg aatgtggacc ccacaccttg cctataccag
     1201 gaagtcccac agtgttcaca tccgggctgc cagcatttgt gtctagtcct actttaccgg
     1261 tggctcccat tccttcaccc gctcccgcaa cacctttacc tccaccggca ctcttacccc
     1321 ccgtaaccac gtcttcctcc ccaatccctc catcccatcc tgtgtctccg gggaccacgg
     1381 atactcattc tccatctcct gcattgccac ccacgcagtc tccagagtct tctcaaaggc
     1441 caccgctttc aagtcctaca ggaaggccag actcttcaac acctatgcgt ccgccaccct
     1501 cgcagcagac tacacctcca cactcaccca cgactcctcc acccgagcct ccctccaagt
     1561 cgtcaccaga ctctttagct ccgtctaccc tgcgtagcct gagaaaaaga aggctatcgt
     1621 ccccccaagg tccctctaca ctaaacccaa tatgtcagtc gcccccagtc tctcccccta
     1681 gatgtgactt cgccaaccgt agtgtgtacc ccccatgggc cacagagtcc ccgatctacg
     1741 tgggatcatc cagcgatggc gatactccgc cacgccaacc gcctacatct cccatctcca
     1801 taggatcatc atccccgtct gagggatcct ggggtgatga cacagccatg ttggtgctcc
     1861 ttgcggagat tgcagaagaa gcatccaaga atgaaaaaga atgttccgaa aataatcagg
     1921 ctggcgagga taatggggac aacgagatta gcaaggaaag tcaggttgac aaggatgaca
     1981 atgacaataa ggatgatgag gaggagcagg agacagatga ggaggacgag gaggatgacg
     2041 aggaggatga cgaggaggat gacgaggagg atgacgagga ggatgacgag gaggatgacg
     2101 aggaggatga cgaggaggat gacgaggagg aggacgagga ggaggacgag gaggaggagg
     2161 acgaggagga ggaggaggag gacgaggagg atgacgatga tgaggacaat gaggacgagg
     2221 aggaggacaa gaaggaggac gaggaggacg ggggcgatgg aaacaaaacg ttgagcatcc
     2281 aaagttcaca acagcagcag gagccacagc agcaggagcc acaacagcag gagccacagc
     2341 agcaggagcc acagcagcag gagcccctgc aggagccaca gcagcaggag ccacaacagc
     2401 aggagccaca acagcaggag ccacaacagc aggagccaca acagcaggag ccacaacagc
     2461 aggagccaca gcagcaggat gagcagcagc aggatgagca gcagcaggat gagcagcagc
     2521 aggatgagca gcagcaggat gagcaggagc agcaggatga gcagcagcag gatgagcagc
     2581 agcaggatga gcagcagcag caggatgaac aggagcagca ggaggagcag gagcagcagg
     2641 aggagcagga gcagcaggag gagcaggagc aggagttaga ggagcaggag caggagttag
     2701 aggagcagga gcaggagtta gaggagcagg agcaggagtt agaggagcag gagcaggagt
     2761 tagaggagca ggagcaggag ttagaggagc aggagcagga gttagaggag caggagcagg
     2821 agttagagga gcaggagcag gagttagagg agcaggagca ggagttagag gagcaggagc
     2881 aggagttaga ggagcaggag caggagttag aggagcagga gcaggagtta gaggagcagg
     2941 agcaggagtt agaggagcag gagcaggagt tagaggagca ggagcaggag ttagaggagc
     3001 aggagcagga gttagaggag caggagcagg agttagagga gcaggagcag gagttagagg
     3061 agcaggagca ggagttagag gagcaggagc aggagcagga gttagaggag gtggaagagc
     3121 aagagcagga gcaggaagag caggaattag aggaggtgga ggagcaagag caggagcagg
     3181 aggagcagga ggagcaggag ttagaggagg tggaagagca ggaagagcag gagttagagg
     3241 aggtggaaga gcaggaagag caggagttag aggaggtgga agagcaggag cagcaggggg
     3301 tggaacagca ggagcaggag acggtggaag agcccataat cttgcacggg tcgtcatccg
     3361 aggacgaaat ggaagtggat taccctgttg ttagcacaca tgaacaaatt gccagtagcc
     3421 caccaggaga taatacacca gacgatgacc cacaacctgg cccatctcgc gaataccgct
     3481 atgtactcag aacatcacca ccccacagac ctggagttcg tatgaggcgc gttccagtta
     3541 cccacccaaa aaagccacat ccaagatacc aacaaccacc ggtcccttac agacagatag
     3601 atgattgtcc tgcgaaagct aggccacaac acatctttta tagacgcttt ttgggaaagg
     3661 atggaagacg agatccaaag tgtcaatgga agtttgcagt gattttttgg ggcaatgacc
     3721 catacggact taaaaaatta tctcaggcct tccagtttgg aggagtaaag gcaggccccg
     3781 tgtcctgctt gccccaccct ggaccagacc agtcgcccat aacttattgt gtatatgtgt
     3841 attgtcagaa caaagacaca agtaagaaag tacaaatggc ccgcctagcc tgggaagcta
     3901 gtcaccccct ggcaggaaac ctacaatctt ccatagttaa gtttaaaaag cccctgccat
     3961 taacccagcc aggggaaaac caaggtcctg gggactctcc acaggaaatg acactcgagg
     4021 gaggcggtgg atggagccac ccccagttcg agaagggcgg cggcagcggc ggcggcagcg
     4081 gcggcggcag ctggagccac ccccagttcg agaaggggcc cgtttaaacc cgctgatcag
     4141 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4201 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4261 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4321 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4381 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4441 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4501 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4561 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4621 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4681 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4741 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4801 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4861 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4921 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4981 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5041 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5101 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5161 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5221 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     5281 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     5341 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     5401 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     5461 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     5521 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     5581 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     5641 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     5701 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5761 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5821 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5881 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5941 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6001 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6061 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6121 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6181 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6241 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6301 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6541 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6781 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6901 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6961 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7021 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7081 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7141 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7201 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7261 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7321 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7381 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7441 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7501 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7561 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7621 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7681 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7741 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7801 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7861 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7921 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7981 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8041 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8101 cgcacatttc cccgaaaagt gccacctgac gtc
//
