LOCUS       sequence-391760-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV protein ORF18 fused at the C-terminal to 
            TurboID-3xHA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  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 6890)
  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_ORF18_TurboID_3xHA
FEATURES             Location/Qualifiers
     source          1..6890
                     /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             1802..2758
                     /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             2786..2812
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2816..2842
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2846..2872
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2898..2915)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2904..3128
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3174..3602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3471..3492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3616..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3858..3877
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3993..4040
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4059..4433
                     /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    4563..4684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4600..4619)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4654..4673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4733..4749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4733..4749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4746..4768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4757..4773
                     /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(4781..4787,4788..4805,4806..4811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4826..4847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4964..4981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5135..5723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5215..5234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5894..6685,6686..6754))
                     /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(6755..6859)
                     /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 atgctcggaa aatacgtgtg
     1021 tgagaccgag ccgctgagcc cggggcttag gaggctcatg tggcgctttt tgcaaaataa
     1081 gaatttaaat acattccacg cccaagagct gcgttttatt catttggttc tctgcaggat
     1141 gtacaatttc ggtctaaatg tgtacctgtt aagggaggct actgccaatg ccgggaccta
     1201 cgacgaggtg gtcctgggac gcaaggttcc tgcggaggtg tggaagctcg tgtacgatgg
     1261 gctcgaggag atgggcgtgt caagtgagat gctgctgtgt gaggcatacc gggacagcct
     1321 ctggatgcac ttgaacgata aggtggggct cttgaggggc ctggcgaatt atctgtttca
     1381 ccggctaggg gtcacccacg acgttcgcat cgccccggaa aacctggtgg acggaaactt
     1441 tttgtttaat ctgggaagtg tgctcccctg caggctgctc cttgcggcgg gctactgcct
     1501 cgccttttgg ggcagcgatg aacacgaacg ctgggtgcgc ttcttcgccc agaagctttt
     1561 catttgctac ctgatagtct ccgggcgtct tatgacacag aggtctctgc tagtttgggc
     1621 cagcgaaacg ggctatcccg gtccggtgga ggcagtctgt cgcgacatcc gctccatgta
     1681 cggcatacga acgtatgcgg tctcgggtta tcttccggct ccgtccgaag cgcagctggc
     1741 ctaccttggt gcgtttaaca acaacgcggt tgggtcaggc ggctctggtg gctccggagc
     1801 caaagacaat actgtgcctc tgaagctgat cgctctcctg gctaatggcg agttccatag
     1861 tggcgaacag ctgggagaaa ccctgggcat gtccagggcc gctatcaaca agcacattca
     1921 gactctgcgc gactggggcg tggacgtgtt caccgtgccc ggaaagggct actctctgcc
     1981 cgagcctatc ccgctgctga acgctaaaca gattctggga cagctggacg gcgggagcgt
     2041 ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct
     2101 gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag
     2161 gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg
     2221 gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg
     2281 aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag
     2341 aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt
     2401 cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg
     2461 ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctacgct
     2521 gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct
     2581 gccacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga
     2641 caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca
     2701 ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagtc
     2761 gcggccgctc gagggaggcg gtggataccc gtatgatgtt ccggattacg ctggctatcc
     2821 ctacgacgtg cccgactatg ccgggtaccc ctatgacgtc ccagactacg cagggcccgt
     2881 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2941 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3001 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3061 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3121 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3181 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3241 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3301 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3361 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3421 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3481 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3541 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3601 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3961 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     4021 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     4081 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     4141 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     4201 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     4261 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     4321 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     4381 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     4441 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4501 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4561 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4621 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4681 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4741 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4801 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4921 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4981 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5041 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5101 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5161 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5221 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5701 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5821 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5881 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5941 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6001 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6061 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6121 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6181 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6241 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6301 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6361 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6421 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6481 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6541 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6601 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6661 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6721 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6781 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6841 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
