LOCUS       sequence-267499-        6209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally strep tagged Kaposi's Sarcoma Associated 
            Herpesvirus (KSHV) ORF29b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6209)
  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 6209)
  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-ORF29b-2xCSTREP
FEATURES             Location/Qualifiers
     source          1..6209
                     /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             2108..2131
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2168..2191
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2217..2234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2223..2447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2493..2921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2580..2599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2790..2811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2930..2950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2935..3264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3115..3250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3177..3196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3312..3359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3378..3752
                     /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    3882..4003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3919..3938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3973..3992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4052..4068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4052..4068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4065..4087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4076..4092
                     /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(4100..4106,4107..4124,4125..4130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4145..4166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4283..4300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4454..5042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4534..4553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5213..6004,6005..6073))
                     /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     5836..5855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6074..6178)
                     /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 attcatgctt cagaaagacg ccaagctgat atttatatca tccgtgaact cgtcagaccg
     1081 ctccacgagt ttcctgctta acctcaggaa cgcccaggaa aagatgctga atgtggtcag
     1141 ttacgtgtgt gcggaccacc gcgaagattt ccacctgcaa gacgcactag tgtcctgtcc
     1201 ttgttacaga ctgcacattc cgacgtacat caccatcgac gaatccatca aaaccaccac
     1261 caacctcttt atggaggggg catttgacac cgaactaatg ggcgagggag cagcgtcgtc
     1321 aaatgctacg ctttaccgcg tggtgggtga cgcagcgctg acacagtttg acatgtgtcg
     1381 ggtagacacc accgcccagc aggttcagaa gtgccttgga aaacagctgt ttgtttacat
     1441 cgaccccgcg tatacgaaca acacggaggc gtccggtact ggcgtgggcg ccgttgtcac
     1501 gagtactcag actcccacca gaagcctcat attgggcatg gagcatttct tcctgcgcga
     1561 cctcactggc gcagctgctt acgagatagc gtcctgcgca tgcacgatga ttaaggcgat
     1621 cgctgtgctc caccccacaa ttgagcgcgt gaacgcggcg gtcgaaggca acagcagcca
     1681 agattctggg gtggccattg caaccgtcct taacgaaata tgcccgctcc ccatacattt
     1741 tctacactat actgacaaga gcagcgccct gcagtggcca atttacatgt tgggaggcga
     1801 gaaatcctcc gcgtttgaga cattcatcta cgctctgaac tccggcaccc tgagcgccag
     1861 ccagacggtg gtgtccaaca ccatcaaaat atcatttgac ccggtgacct acctggtaga
     1921 acaggtccgc gcgatcaagt gcgtcccgct tagggatgga gggcagtcat acagcgccaa
     1981 gcaaaagcac atgtcggacg acttacttgt ggcagttgtc atggcccatt ttatggctac
     2041 cgatgataga cacatgtaca agcccatatc cccacaatca gggcggccgc tcgagggagg
     2101 cggtggatgg agccaccccc agttcgagaa gggcggcggc agcggcggcg gcagcggcgg
     2161 cggcagctgg agccaccccc agttcgagaa ggggcccgtt taaacccgct gatcagcctc
     2221 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2281 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2341 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2401 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2461 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2521 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2581 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2641 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2701 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2761 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2821 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2881 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2941 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3001 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3061 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3181 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3241 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3301 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     3361 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     3421 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     3481 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     3541 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     3601 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     3661 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     3721 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     3781 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3841 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3901 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3961 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4021 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4081 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4201 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4261 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4321 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4861 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5341 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6061 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6181 catttccccg aaaagtgcca cctgacgtc
//
