LOCUS       sequence-268949-        5921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF46.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5921)
  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 5921)
  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-ORF46-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..5921
                     /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             1820..1843
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1880..1903
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1929..1946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1935..2159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2205..2633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2292..2311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2502..2523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2642..2662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2647..2976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2827..2962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2889..2908
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3024..3071
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3090..3464
                     /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    3594..3715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3631..3650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3685..3704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3764..3780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3777..3799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3788..3804
                     /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(3812..3818,3819..3836,3837..3842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3857..3878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3995..4012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4166..4754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4246..4265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4925..5716,5717..5785))
                     /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     5548..5567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5786..5890)
                     /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 attcatggac gcatggttgc aacagacggt ctttaggggc accctatcca tcagtcaggg
     1081 ggtggacgac cgggatctgt tactggcacc taagtggatt tcctttctga gcctctcatc
     1141 atttctgaaa cagaaactgc tctcgctgct cagacagatt cgggaactta ggctaaccac
     1201 cacagtgtat cccccacagg acaagctgat gtggtggtcc cactgctgcg atccagagga
     1261 tattaaagtg gtgatcttag gccaggaccc gtaccacaag ggccaagcta ctggcctggc
     1321 gtttagtgtg gatccgcaat gtcaggttcc acccagtttg agaagcatct ttagagagct
     1381 agaggcttcc gtccccaatt tcagtactcc ttcccacggg tgcctcgaca gctgggctcg
     1441 ccagggtgtg ttgctactaa acacagtttt gacggtggag aaggggaggg ccggctcaca
     1501 cgagggactt ggctgggatt ggttcacgag tttcatcatc agtagcatat cctcaaagtt
     1561 agaacattgc gtttttctcc tgtgggggcg caaggccatt gacagaactc cgctcataaa
     1621 cgcacagaaa cacctggtgc ttacggccca gcatccatct ccgctggcct ctcttggtgg
     1681 ccgacactcg cgatggcctc ggttccaggg ctgtaatcac tttaacctag ccaacgacta
     1741 tttgactcgc caccggcgtg agactgtgga ctggggcctg ttggagcagt cagggcggcc
     1801 gctcgaggga ggcggtggat ggagccaccc ccagttcgag aagggcggcg gcagcggcgg
     1861 cggcagcggc ggcggcagct ggagccaccc ccagttcgag aaggggcccg tttaaacccg
     1921 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1981 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2041 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2101 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2161 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2221 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2281 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2341 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2401 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2461 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2521 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2581 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2641 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2701 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2761 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2821 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2881 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     2941 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3001 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     3061 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     3121 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     3181 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     3241 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     3301 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     3361 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     3421 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     3481 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3541 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3601 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3661 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3721 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4081 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4741 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4801 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4861 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4921 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4981 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5041 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5101 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5161 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5221 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5281 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5341 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5401 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5461 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5521 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5581 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5641 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5701 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5761 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5821 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5881 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
