LOCUS       sequence-291859-        7485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF44.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  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 7485)
  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-ORF44-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..7485
                     /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             3384..3407
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3444..3467
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3493..3510)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3499..3723
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3769..4197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3856..3875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4066..4087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4206..4226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4211..4540
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4391..4526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4453..4472
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4588..4635
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4654..5028
                     /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    5158..5279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5195..5214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5249..5268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5328..5344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5328..5344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5341..5363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5352..5368
                     /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(5376..5382,5383..5400,5401..5406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5421..5442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5559..5576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5730..6318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5810..5829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6489..7280,7281..7349))
                     /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     7112..7131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7350..7454)
                     /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 cttggtacca tggacagctc ggaagggtgc actgatatgg
     1021 acgagcccag ccccgggttc atcctcaaca tgacatcgga tgccaaagtc aggagcgtag
     1081 tggaacagat tgacaggttg tcaaatatca ctacctcgcc cccggagatg ggctggtatg
     1141 acctagagtt cgatccactg gaagacgaag gcccctttct gccgttttcg gcatacgtaa
     1201 taacggggac tgcaggagcg gggaaaagca ccagcgtatc cgccctacat cagaatctca
     1261 actgcctaat tacgggggct acagtggtag cggcacagaa tctttccagg gctttaaagt
     1321 cctactgtcc cactatatac cacgccttcg gattcaagag cagacacatt aatatctgcc
     1381 agaggaaagt gcccaaggta actcagtcct ccatcgagca actccagaga tacgagctgg
     1441 ctaggtactg gccaactgtc accgatatta ttcgagaatt tatgcgcaag aaacaaaagg
     1501 ggcagtatag ctccctctct caaagcgctt tcagactcct ttgccgtatg ggtggagcca
     1561 atttgtggac gagtaacatt atcgtgatag acgaagctgg aaccctctcg tcccatattt
     1621 tgacggccgt ggtgttcttc tattggtttt acaacagttg gctggacacc ccgctataca
     1681 gaaatggtgc cgtgccttgc atagtctgcg tggggtctcc cacccagacg gacgcctttc
     1741 agtcggtctt caaccacacg cagcagagaa acgagatatc tgcctgtgat aatgtgctca
     1801 ccttcctatt gggaaaacgt gaggttgcag attatattag gctggacgag aattgggccc
     1861 tatttataaa caataagcgc tgtacggatc cccagtttgg tcacttgctg aagaccttag
     1921 aatataatct agacatatca ccagagttaa tggactatat agataggttt gtggttccga
     1981 agagtaagat tctggacccg ctcgagtatg cagggtggac aagactcttc atctcacacc
     2041 aggaggtgaa gtcttttctg gcaacgctgc acacctgcct gtcgagtaat aaggatgctg
     2101 tgtccacaaa gcttttcacc tgcccagtgg tctgtgaggt gtttacagag ccatttgagg
     2161 agtacaaacg ggcggtaggc ctcacacaca tgactcccat agaatgggta acaaaaaatc
     2221 ttttcaggct aagtaactac tcgcagtttg ctgatcagga catggctgtg gttgggacct
     2281 atatcacaga cgcgtccaca cagatcacct tcgccactaa atttgtcaaa aacagctatg
     2341 ctacccttac tggaaagacc aaaaaatgta tatgcgggtt tcacgggtca taccaaagat
     2401 tcaagtccat cctagacggg gagctattta tcgaaagtca ttcgcacgat aaccccgctt
     2461 atgtgtacag tttccttagt accctgctat ataatgccat gtactcattt tacgcgcacg
     2521 gggtgaagca ggggcatgaa gaattcctca gggacctcag ggaactgccg gtgtctcaag
     2581 agctgatctc tgagatgagc tccgaggacg ttctggggca ggagggggac acagatgcct
     2641 tctacctcac cgccagcctc ccaccatccc ccacccacgc ggctcttcca acactggtgg
     2701 cctattactc cggggccaag gaactattct gcaacaggct ggccctggca cgccgacact
     2761 ttggtgacga gttcctccac tccgattttt caacgtttac ggtgaacatc gtggtgcgag
     2821 atggcgtgga ctttgtgtcc acttcccccg ggctccacgg tctagtggca tacgcatcca
     2881 ctatagacac ctatataatc cagggatata cgttcctccc agtgagattc ggccgtccag
     2941 gaggacagcg cctcagcgag gacctgcgca gaaagatgcc ctccatagtt gtccaggact
     3001 catcggggtt cattgcctgc ctggaaaata acgtcaccaa gatgacagag accctcgaag
     3061 gtggcgacgt gtttaacata tgttgtgcag gggactacgg tatcagttct aatctggcta
     3121 tgaccatagt gaaggcacag ggggtttcac taagtagggt ggccatatcg ttcggcaacc
     3181 accgcaatat cagagccagt ctagtgtatg tgggtgtatc cagggccatc gacgctcgtt
     3241 acctggtaat ggacagtaat ccccttaagc taatggaccg cggtgacgcc cagtccccat
     3301 cctcaaagta catcatcaaa gccctatgca accccaagac tactctgatc tacggagggc
     3361 ggccgctcga gggaggcggt ggatggagcc acccccagtt cgagaagggc ggcggcagcg
     3421 gcggcggcag cggcggcggc agctggagcc acccccagtt cgagaagggg cccgtttaaa
     3481 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3541 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3601 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3661 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3721 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3961 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4201 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4261 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4321 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4381 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4441 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4501 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4561 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     4621 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     4681 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     4741 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     4801 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     4861 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     4921 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     4981 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     5041 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5101 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5161 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5221 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5281 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5341 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5401 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5461 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5521 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5581 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5641 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5701 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5761 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5821 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5881 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5941 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6001 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6061 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6121 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6181 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6241 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6301 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6361 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6421 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6481 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6721 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6781 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6901 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7021 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7141 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7201 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7321 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7381 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7441 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
