LOCUS       sequence-267627-        5622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF65.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  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 5622)
  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-ORF65-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..5622
                     /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             1521..1544
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1581..1604
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1630..1647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1636..1860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1906..2334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1993..2012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2203..2224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2343..2363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2348..2677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2528..2663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2590..2609
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2725..2772
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2791..3165
                     /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    3295..3416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3332..3351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3386..3405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3465..3481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3465..3481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3478..3500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3489..3505
                     /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(3513..3519,3520..3537,3538..3543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3558..3579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3696..3713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3867..4455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3947..3966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4626..5417,5418..5486))
                     /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     5249..5268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5487..5591)
                     /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 cttggtacca tgtccaactt taaggtgaga gaccccgtga
     1021 tccaggagcg actggatcat gactacgctc accatcccct ggtggctcgc atgaataccc
     1081 tggatcaggg aaatatgtcg caggccgaat accttgtgca aaagagacac tatctcgtgt
     1141 tcttaattgc ccatcattat tatgaggcct atctgaggag gatgggtgga attcaaaggc
     1201 gggatcatct gcagaccctt agggatcaga aaccgcgcga gcgcgctgac cgagtttccg
     1261 cggcgtcggc ttacgacgcg gggacgttca ccgtgccttc gaggccaggc cctgcatccg
     1321 gcgccacgcc cgggggccag gactcactgg gggtctcggg aagcagtata accaccctgt
     1381 cctctggtcc ccattcattg tcgccggcgt cagacattct cacaaccctc tcatccacga
     1441 cggaaacggc cgcccccgcg gtggccgacg cgaggaaacc cccctctggc aaaaagaaat
     1501 cgctcgaggg aggcggtgga tggagccacc cccagttcga gaagggcggc ggcagcggcg
     1561 gcggcagcgg cggcggcagc tggagccacc cccagttcga gaaggggccc gtttaaaccc
     1621 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1681 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1741 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1801 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     1861 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     1921 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1981 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2041 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2101 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2161 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2221 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2281 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     2341 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2401 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2461 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2521 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2581 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     2641 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     2701 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     2761 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     2821 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     2881 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     2941 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     3001 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     3061 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     3121 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     3181 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3241 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3421 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5581 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
