LOCUS       sequence-391735-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV protein ORF66 fused at the C-terminal to 
            TurboID-3xHA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  AUTHORS   McCollum CO, Didychuk AL, Liu D, Murray-Nerger LA, Cristea IM, 
            Glaunsinger BA
  TITLE     The viral packaging motor potentiates Kaposi's sarcoma-associated 
            herpesvirus gene expression late in infection.
  JOURNAL   PLoS Pathog. 2023 Apr 17;19(4):e1011163. doi: 
            10.1371/journal.ppat.1011163. eCollection 2023 Apr.
  PUBMED    37068108
REFERENCE   2  (bases 1 to 7406)
  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"; journalName: "PLoS 
            Pathog."; date: "2023-04-17"; volume: "19(4):e1011163. doi"; pages: 
            " 10.1371/journal.ppat.1011163. eCollection 2023 Apr"
COMMENT     Sequence Label: pCDNA4.TO_ORF66_TurboID_3xHA
FEATURES             Location/Qualifiers
     source          1..7406
                     /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             2318..3274
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             3302..3328
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3332..3358
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3362..3388
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3414..3431)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3420..3644
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3690..4118
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3777..3796)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3987..4008
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4127..4147)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4132..4461
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4312..4447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4374..4393
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4509..4556
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4575..4949
                     /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    5079..5200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5116..5135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5170..5189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5249..5265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5249..5265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5262..5284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5273..5289
                     /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(5297..5303,5304..5321,5322..5327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5342..5363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5480..5497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5651..6239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5731..5750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6410..7201,7202..7270))
                     /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"
     promoter        complement(7271..7375)
                     /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 atggccctgg atcagcgctg
     1021 ggatcgcttt cttgtgtctt ggtttggttt ggatgaggcc cagctgacgg cccaccgcgt
     1081 gttcgagggc gagaatggcg tgccagtgga ggaatatgtg gcttttgtca tattcgggga
     1141 gcggggtttc cagggaaaca tgccctcctg ggccaggcac ctgttggacc gcccgtcgct
     1201 ggcgcaggcg atcgccgtgt tgcgcgctgg ctcagacacc gtcgccaaac aggcgcagat
     1261 ttgtgcggcg cagcagctgc ttggcgccca cgtatgggtt gtggtgacgc tgtcccgtgc
     1321 ccaggccgcc gaccatgcta gggcgatacc gcgccacgta tgggcaaaat acctgtcgct
     1381 tccattttcc aaggcctgtg cccaactctg caagctgttg gcgctttgct ctcgctttcc
     1441 gttagtgacc tgctgttcga agcccccacc gtcgctaccc tggctaagga agaagtggca
     1501 tgggccgttg ccgcgtcgtc ccctcctgga ggtgccctcg cccacccgtc gcggtgtggc
     1561 tgcgaccgga gacggtaacg ggctcgggat aggggcggca gataccggcc tccgggaggc
     1621 cttggagcgc gtcgccccta cggttccctg cggtaacccg ttcgacgcca tgcttgggtc
     1681 actgtgtttt ttgagcctta ttaagtcccg ccacgtagtg ctgcccgcct gcgagcagga
     1741 gggacctggt ctcgtgcgga accttggcag gaggcttctg gcatacaatg ttctctcccc
     1801 ctgcgtgtcc atacccgtca tctgttcgcg cgtggcccgc gcggcccttg ccaagcgggc
     1861 gcgttgcgct cgggcggttg tgtgcatgga gtgtggacac tgtcttaact ttggcagggg
     1921 caagtttcat actgtcaatt ttcctcccac caacgtgttt ttcagcaggg acaggaaaga
     1981 gaagcagttc accatctgtg cgaccacggg gaggatctac tgttcttact gtggcagcga
     2041 acatatgagg gtgtatcccc tgtgcgatat taccggacgc gggaccctag cacgcgtcgt
     2101 aatcagggct gtcctagcta acaacgcggc ccttgccatt cgagacctag atcaaactgt
     2161 cagttttgta gtgccttgcc ttgggacgcc cgactgcgag gctgccctat taaagcaccg
     2221 tgacgtgcgc ggactgcttc agctcacctc acagctgctg gagttctgtt gcgggaagtg
     2281 ttcctccggg tcaggcggct ctggtggctc cggagccaaa gacaatactg tgcctctgaa
     2341 gctgatcgct ctcctggcta atggcgagtt ccatagtggc gaacagctgg gagaaaccct
     2401 gggcatgtcc agggccgcta tcaacaagca cattcagact ctgcgcgact ggggcgtgga
     2461 cgtgttcacc gtgcccggaa agggctactc tctgcccgag cctatcccgc tgctgaacgc
     2521 taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg tggtcgactc
     2581 caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg cttgcattgc
     2641 agaatatcag caggcaggga gaggaagcag agggaggaaa tggttctctc cttttggagc
     2701 taacctgtac ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa tcggcctggg
     2761 cccggtcatc ggaattgtca tggcagaagc gctgcgaaag ctgggagcag acaaggtgcg
     2821 agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca tcctggtgga
     2881 gctggccgga ataacaggcg atgctgcaca gatcgtcatt ggcgccggga ttaacgtggc
     2941 tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc aggaagcagg
     3001 gattaacctg gacaggaata ctctggccgc tacgctgatc cgagagctgc gggcagccct
     3061 ggaactgttc gagcaggaag gcctggctcc atatctgcca cggtgggaga agctggataa
     3121 cttcatcaat agacccgtga agctgatcat tggggacaaa gagattttcg ggattagccg
     3181 ggggattgat aaacagggag ccctgctgct ggaacaggac ggagttatca aaccctggat
     3241 gggcggagaa atcagtctgc ggtctgccga aaagtcgcgg ccgctcgagg gaggcggtgg
     3301 atacccgtat gatgttccgg attacgctgg ctatccctac gacgtgcccg actatgccgg
     3361 gtacccctat gacgtcccag actacgcagg gcccgtttaa acccgctgat cagcctcgac
     3421 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3481 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3541 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3601 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3661 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3721 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3781 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3841 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3901 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3961 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4021 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4081 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4141 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4201 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4261 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4321 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4381 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4441 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4501 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4561 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     4621 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     4681 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     4741 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     4801 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     4861 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     4921 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     4981 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5041 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5161 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5221 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5281 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5341 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5401 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5461 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5521 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5581 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5641 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5701 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5821 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6001 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6061 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6121 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6181 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6241 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6301 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6361 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6421 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6481 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6541 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6601 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6661 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6721 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6781 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6841 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6901 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6961 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7021 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7081 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7141 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7201 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7261 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7321 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7381 ttccccgaaa agtgccacct gacgtc
//
