LOCUS       sequence-320632-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal 2x-Strep-tagged ORF68 K450A/R451A in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  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 6516)
  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-ORF68 K450A/R451A
FEATURES             Location/Qualifiers
     source          1..6516
                     /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"
     regulatory      1357..1366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2523..2540)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2529..2753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2799..3227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2886..2905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3096..3117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3236..3256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3241..3570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3421..3556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3483..3502
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3618..3665
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3684..4058
                     /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    4188..4309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4225..4244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4279..4298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4358..4374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4358..4374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4371..4393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4382..4398
                     /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(4406..4412,4413..4430,4431..4436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4451..4472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4589..4606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4760..5348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4840..4859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5519..6310,6311..6379))
                     /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     6142..6161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6380..6484)
                     /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 ccatgtttgt tccctggcaa ctcggtacaa ttacccgtca ccgagatgag ctccaaaaac
     1141 tactggcagc ctccctgctc ccggagcacc cggaggagag cctcggtaac cccataatga
     1201 cacagattca ccagtcgctc caaccatctt ccccctgcag ggtctgtcag ctcctatttt
     1261 ctctggtccg cgattcgtcc acccccatgg gtttcttcga ggactatgcc tgcctctgct
     1321 tcttctgtct atacgcccca cactgctgga cctcgaccat ggcggcagcg gcagacctgt
     1381 gcgagatcat gcatctgcac tttccagaag aggaggcgac atacgggcta ttcggaccgg
     1441 gtcgccttat gggtatcgac ttgcagctgc acttctttgt tcaaaagtgc tttaagacca
     1501 ccgccgccga aaaaatactg ggaatatcca acctgcaatt tttaaaatca gaattcatcc
     1561 ggggcatgct cacaggcacc atcacctgca acttctgctt caaaacgtcc tggcccagga
     1621 cagacaagga ggaggccacc ggccccaccc catgctgcca gattacagac accaccaccg
     1681 cacccgcgag cggcataccg gaactagccc gggccacatt ctgcggcgca agtcgcccca
     1741 caaagcccag cctacttccc gcgctaatag atatctggtc cacgagctca gagctccttg
     1801 acgagccgcg ccctcgactg atcgcaagcg acatgagtga actcaaatcc gtggtcgcat
     1861 cccacgatcc gttcttctct cccccgcttc aggcagacac ctcccagggt ccatgtctaa
     1921 tgcacccaac cctggggcta cgatacaaaa acgggactgc atccgtctgc ctcctctgcg
     1981 agtgccttgc ggcacaccca gaggcaccca aggcgctgca gacccttcag tgcgaggtaa
     2041 tgggccatat agaaaacaac gtaaagctgg tagacagaat tgcctttgtg ttggacaacc
     2101 cattcgccat gccatatgtc tcagatccgc tacttagaga gctgatccgg ggctgtaccc
     2161 cacaggaaat tcacaagcac ctgttctgcg acccgctgtg cgccctcaat gctaaggtgg
     2221 tgtcagagga cgtactattc cgcctgccca gggagcagga gtataaaaag ctcagggcat
     2281 ccgcggccgc cggacagctc ctcgatgcca acaccctgtt cgactgcgag gtcgtgcaga
     2341 ctttggtctt tctctttaag ggtctccaaa acgccagggt ggggaaaacc acctcactag
     2401 acattattcg ggagctaacc gcacaactag ccgcccaccg cctagacctg gcccacccct
     2461 cacagacgtc acacttgtac gcttaatcga gtctagaggg cccgtttaaa cccgctgatc
     2521 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2581 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2641 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2701 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2761 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2821 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2881 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2941 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3001 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3061 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3121 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3181 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3241 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3421 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3481 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3541 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3601 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     3661 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     3721 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     3781 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     3841 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     3901 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     3961 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     4021 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
     4081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6481 cgcgcacatt tccccgaaaa gtgccacctg acgtcg
//
