LOCUS       sequence-391648-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV protein ORF34 fused at the N-terminal to 
            3xHA-TurboID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  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 7079)
  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_3xHA_TurboID_ORF34
FEATURES             Location/Qualifiers
     source          1..7079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    819..837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    840..858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     868..892
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             986..1012
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1016..1042
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1046..1072
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1082..2038
                     /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"
     primer_bind     complement(3086..3103)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3092..3316
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3362..3790
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3449..3468)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3659..3680
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3799..3819)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3804..4133
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3984..4119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4046..4065
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4181..4228
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4247..4621
                     /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    4751..4872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4788..4807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4842..4861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4921..4937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4921..4937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4934..4956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4945..4961
                     /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(4969..4975,4976..4993,4994..4999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5014..5035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5152..5169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5323..5911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5403..5422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6082..6873,6874..6942))
                     /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(6943..7047)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct
      841 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag
      901 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
      961 tctagcgttt aaacttaagc ttatgtaccc gtatgatgtt ccggattacg ctggctatcc
     1021 ctacgacgtg cccgactatg ccgggtaccc ctatgacgtc ccagactacg caggggcggc
     1081 caaagacaat actgtgcctc tgaagctgat cgctctcctg gctaatggcg agttccatag
     1141 tggcgaacag ctgggagaaa ccctgggcat gtccagggcc gctatcaaca agcacattca
     1201 gactctgcgc gactggggcg tggacgtgtt caccgtgccc ggaaagggct actctctgcc
     1261 cgagcctatc ccgctgctga acgctaaaca gattctggga cagctggacg gcgggagcgt
     1321 ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct
     1381 gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag
     1441 gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg
     1501 gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg
     1561 aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag
     1621 aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt
     1681 cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg
     1741 ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctacgct
     1801 gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct
     1861 gccacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga
     1921 caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca
     1981 ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaaggg
     2041 gtcaggcggc tctggtggct ccggagcctt tgctttgagc tcgctcgtgt ccgagggtga
     2101 cccggaggtg accagtaggt acgtcaaggg cgtacaactt gccctggacc ttagcgagaa
     2161 cacacctgga caatttaagt tgatagaaac tcccctgaac agcttcctct tggtttccaa
     2221 cgtgatgccc gaggtccagc caatctgcag tggccggccg gccttgcggc cagactttag
     2281 taatctccac ttgcctagac tggagaagct ccagagagtc ctcgggcagg gtttcggggc
     2341 ggcgggtgag gaaatcgcac tggacccgtc tcacgtagaa acacacgaaa agggccaggt
     2401 gttctacaac cactatgcta ccgaggagtg gacgtgggct ttgactctga ataaggatgc
     2461 gctccttcgg gaggctgtag atggcctgtg tgaccccgga acttggaagg gtcttcttcc
     2521 tgacgacccc cttccgttgc tatggctgct gttcaacgga cccgcctctt tttgtcgggc
     2581 cgactgttgc ctgtacaagc agcactgcgg ttacccgggc ccggtgctac ttccaggtca
     2641 catgtacgct cccaaacggg atcttttgtc gttcgttaat catgccctga agtacaccaa
     2701 gtttctatac ggagattttt ccgggacatg ggcggcggct tgccgcccgc cattcgctac
     2761 ttctcggata caaagggtag tgagtcagat gaaaatcata gatgcttccg acacttacat
     2821 ttcccacacc tgcctcttgt gtcacatata tcagcaaaat agcataattg cgggtcaggg
     2881 gacccacgtg ggtggaatcc tactgttgag tggaaaaggg acccagtata taacaggcaa
     2941 tgttcagacc caaaggtgtc caactacggg cgactatcta atcatcccat cgtatgacat
     3001 accggcgatc atcaccatga tcaaggagaa tggactcaac caactctaat cgagtctaga
     3061 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3121 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3181 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3241 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3301 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3361 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3421 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3481 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3541 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3601 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3661 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3721 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3781 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3961 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4021 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4081 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4141 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4201 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     4261 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     4321 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     4381 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     4441 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     4501 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     4561 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     4621 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4681 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4741 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4801 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4861 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4921 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4981 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5041 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5101 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5161 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5581 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5761 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5821 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5881 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5941 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6001 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6061 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7021 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcg
//
