LOCUS       sequence-320640-        6426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal 2x-Strep-tagged mu68 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6426)
  AUTHORS   Didychuk AL, Gates SN, Gardner MR, Strong LM, Martin A, Glaunsinger 
            BA
  TITLE     A pentameric protein ring with novel architecture is required for 
            herpesviral packaging.
  JOURNAL   Elife. 2021 Feb 8;10. pii: 62261. doi: 10.7554/eLife.62261.
  PUBMED    33554858
REFERENCE   2  (bases 1 to 6426)
  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: "Elife."; 
            date: "2021-02-8"; volume: "10. pii: 62261. doi"; pages: " 
            10.7554/eLife.62261"
COMMENT     Sequence Label: pcDNA4/TO-2xStrep-mu68
FEATURES             Location/Qualifiers
     source          1..6426
                     /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             990..1013
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1050..1073
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2433..2450)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2439..2663
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2709..3137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2796..2815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3006..3027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3146..3166)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3151..3480
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3331..3466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3393..3412
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3528..3575
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3594..3968
                     /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    4098..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4135..4154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4189..4208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4268..4284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4268..4284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4281..4303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4292..4308
                     /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(4316..4322,4323..4340,4341..4346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4361..4382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4499..4516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4670..5258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4750..4769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5429..6220,6221..6289))
                     /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     6052..6071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6290..6394)
                     /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 ttaccaatgt ggagccaccc ccagttcgag aagggcggcg
     1021 gcagcggcgg cggcagcggc ggcggcagct ggagccaccc ccagttcgag aagggggcgg
     1081 ccatgtacgt accatggtct ctagagacag tcacctccca cgattcaatc atgcagatgt
     1141 tactttctaa atcctaccta cctgccacac cagaaacagg ctttcatgat cccacactca
     1201 ctcatatctc aagcacgttg ttgcccactt ccaagtgcaa gatatgtaac atgcttttcg
     1261 aagctgtgct ggaacacaat ccccctgatt ctttttttag tgagtatgcg accctctgct
     1321 tctacgtgct gcatgccccc gtgacctggt ccgcaaccta cataacaatc tgtgatatgt
     1381 tgcaaatctt ctctaacgcg ggactaaacc atctcattac tgagaaaata aactctagca
     1441 agatcatggg catagacttt taccttcatt tctttgccag gcaatgcttc aggcacacca
     1501 cccccgatca ggtggctcaa ctggaaaacc tgcaccaggt ccgcctgagt ttgttatcta
     1561 ataccatctg taatgtccag tctgctgctc acatacaagg tatctggtcc agctctctgg
     1621 aaacagccca caagaagggg gcccagagtc cagatccccc cactggctgc cctgtcaagg
     1681 ctgtgcccca ccgtgagttc ttttcaaaca ccaacaggcc aagtcttctc aagatattca
     1741 gcgaggcctg gaagcggtcc ctgttactcg accccgatta ctccgagcca gacacacctg
     1801 tagagacggt ggatacctcc caaggaccgt gcctactctc gtttcccttc tccctcaaat
     1861 gtaggggcaa caccagctcc gtgtgcgtgt tgtgcgaatg tctggcgtgt caccccgagg
     1921 cggccaggat cctccaagag atcaaatcca ccatactcac ctgcctggaa aataacacct
     1981 ctttgctaga cagaaccgcc tacaccctga tgatgtacac cacctctaag ctcatagatg
     2041 atccagacct ggctgaaatc atcaccagtc ggggacccaa tgtactacat aagcaccttt
     2101 tctgtgatcc attctgtgcc ataaacagtc tgaaaactga ccctctggtg ctatttaaat
     2161 atcccagcgg acccaaattt caaattttta caaaagccct cgcttctgga aaatatttgg
     2221 cggagaacgc actatttgac cagccagaaa cagagttgtt agtgttggcg ctgaaggcct
     2281 gtcagttggc acagtttcca aaaaaatttg aagcagaagt gttaaaagga atttccaggt
     2341 tgggaactac gctcaccact gtcagtcctg tgcatttctt ttctgtttac acctaatcga
     2401 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2461 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2521 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2581 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2641 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2701 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2761 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2821 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2881 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2941 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3001 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3061 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3121 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3361 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3421 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3481 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     3541 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     3601 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     3661 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     3721 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     3781 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     3841 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     3901 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     3961 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4021 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4081 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4141 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4201 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4261 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4321 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5101 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5401 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5461 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5521 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5581 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5641 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5701 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5761 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5821 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5881 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5941 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6001 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6061 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6121 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6181 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6241 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6301 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6361 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6421 acgtcg
//
