LOCUS       sequence-267575-        6305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF45.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  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 6305)
  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-ORF45-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..6305
                     /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             2276..2299
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2336..2359
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2385..2402)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2391..2615
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2661..3089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2748..2767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2958..2979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3098..3118)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3142..3352
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3164..3360
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3211..3346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3273..3292
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3408..3455
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3474..3848
                     /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    3978..4099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4015..4034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4069..4088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4148..4164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4148..4164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4161..4183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4172..4188
                     /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(4196..4202,4203..4220,4221..4226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4241..4262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4379..4396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4550..5138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4630..4649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5309..6100,6101..6169))
                     /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     5932..5951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6170..6274)
                     /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 attcatggcg atgtttgtga ggacctcgtc tagcacacac gatgaagaga gaatgcttcc
     1081 aattgaagga gcgcctcgca gacgaccccc cgtgaagttc atattcccac ctccacctct
     1141 ttcatcactt ccaggatttg gcaggccgcg cggctatgct ggacccacgg tgatagatat
     1201 gtctgcccca gacgacgtct tcgccgagga cacgccatcg ccgccagcaa cccctctgga
     1261 tctacagata tccccggatc agtcgagcgg cgaatctgaa tatgacgagg atgaggaaga
     1321 tgaagatgaa gaagaaaatg acgatgttca ggaggaagac gagccagagg ggtaccctgc
     1381 agactttttt caacctttat ctcacttgcg cccgaggcct ctggccagac gggcccatac
     1441 gcccaaaccg gtagcagtgg tagcgggccg cgtgcgcagt tcaacggaca cggcggagtc
     1501 cgaggcgtcc atgggatggg ttagtcagga tgacggattt tcccctgctg ggctctcacc
     1561 ttcagacgac gagggggttg ctatcctgga accgatggcg gcatacactg ggaccggggc
     1621 atacggactt tcacctgctt ccagaaatag tgtacctgga acacaaagtt caccatacag
     1681 cgaccctgat gaagggccct cgtggcgccc cctgcgcgcc gcacccaccg cgatcgtcga
     1741 cctgacatcg gactctgata gcgatgacag ttccaactct ccggacgtga acaatgaggc
     1801 cgcgtttacc gacgcgcgcc atttttccca ccagccaccc tcgtccgagg aggacggaga
     1861 agaccaaggg gaagtattga gtcagagaat cgggctcatg gacgtgggcc agaagcgcaa
     1921 aaggcagtct accgcctcct ctggtagcga ggatgtggtg cgctgccaga gacaaccaaa
     1981 cttaagccgc aaagcagtgg cgtccgtgat aattatatcc tcggggagtg acacagacga
     2041 ggagccctcg tccgccgtga gcgtgatcgt gtctccgtcg agcacaaagg gtcacctccc
     2101 aacccaatct cccagtactt ccgcccactc gatttcatca ggaagcacaa ctaccgcggg
     2161 gtccaggtgc agcgacccaa cccgcatcct ggcctccacg ccacccctgt gtggaaacgg
     2221 tgcatataac tggccgtggc tggacggagg gcggccgctc gagggaggcg gtggatggag
     2281 ccacccccag ttcgagaagg gcggcggcag cggcggcggc agcggcggcg gcagctggag
     2341 ccacccccag ttcgagaagg ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2401 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2461 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2521 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2581 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2641 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2701 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2761 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2821 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2881 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2941 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3001 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3061 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3181 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3241 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3301 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3361 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     3421 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     3481 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     3541 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     3601 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     3661 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     3721 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     3781 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     3841 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3901 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3961 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4081 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6301 acgtc
//
