LOCUS       sequence-320335-        6903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal 2x-Strep-tagged UL32 C308A/C309A in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6903)
  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 6903)
  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-UL32 C308A/C309A
FEATURES             Location/Qualifiers
     source          1..6903
                     /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(2910..2927)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2916..3140
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3186..3614
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3273..3292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3483..3504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3623..3643)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3628..3957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3808..3943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3870..3889
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4005..4052
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4071..4445
                     /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    4575..4696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4612..4631)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4666..4685
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4745..4761)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4745..4761)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4758..4780)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4769..4785
                     /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(4793..4799,4800..4817,4818..4823))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4838..4859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4976..4993)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5147..5735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5227..5246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5906..6697,6698..6766))
                     /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     6529..6548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6767..6871)
                     /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 ccatggcaac ttcgcccccc ggggtcctgg cgagcgtcgc ggtgtgcgaa gagtcccccg
     1141 gcagcagctg gaaggcgggg gcgttcgagc gcgcgtatgt ggccttcgat cccagcctcc
     1201 tggctcttaa cgaggctctc tgcgccgagc tcctggcggc cagtcacgtg ataggtgtgc
     1261 cgcccgtcgg tacgctcgac gaggacgtcg ccgccgacgt ggtcaccgcc ccctcaaggg
     1321 cccgcggggg ggcgggcgac ggagggggtt cgggcgggcg cggaggaccc cgcaacccgc
     1381 ccccggatcc ctgtggggag gggcttttgg acaccgggcc cttttccgcg gcggccatcg
     1441 acaccttcgc gctggaccgg ccctgtctgg tctgccgcac catcgagctg tacaagcaaa
     1501 cctaccgcct ctcgccccag tgggtggccg attacgcgtt tctctgtgcc aaatgcctgg
     1561 gggcgcccca ctgcgccgct agcatcttcg tggccgcgtt cgagtttgtg tacgtcatgg
     1621 accgccactt tctgcgcacc aagaaggcga ccctggtcgg ctccttcgca cgctttgccc
     1681 tcaccataaa cgacatccac cggcactttt ttctccactg ctgcttccgt accgacggcg
     1741 gggtgcccgg gcggcatgcc cagaagcaac ccaagccctc gccctccccg ggagccgcca
     1801 aggtgcagta ctccaactac tcgttcctgg cgcagtctgc cacccgggcc ctcatcggaa
     1861 ccttggcctc cgggggcgag gagggggcgg ggtcggcggc gggatccggc acgcagccgt
     1921 cgctcaccac cgcgctgatg aactggaagg attgcgcccg tctcctggac tgcacggagg
     1981 gtcggcgcgg gggcggggac agcgccgcca cgcgcgccgc cgctcgcaac ggggaattcg
     2041 agacggtcgc cggggaccgc gagccggagg agtcgccgga tacgtgggcg tacgcggatc
     2101 tggtcctgct gctcctcgcc ggcacccccg ccgtctggga gtcggggccc cagctgcgcg
     2161 ctgccgcgga ggcccgtcgc gccacggtcc gccagtcctg ggaagcgcat cgcggggcac
     2221 gcacgcgcga cgtggctccg cgctttgcgc agtttaccga acccgatgcc cagcccgacc
     2281 tcgacctggg ccctctgatg gccaccgtgc tgaaacacgg ccgggggcgc gggcgcaccg
     2341 gcggagaatg cctgctgtgt aacctgttac tggtgcgggc ttactggctg gccctgcgta
     2401 gactccgggc ctccgtggtc cgctactcgg aaaacaacac gagcctcttc gactgtatcg
     2461 tacccgtcgt cgaccagctc gaggcggacc ccgagacgca gcccggggac gggggccggt
     2521 ttgtgagcct gctgcgggcc gcggggcccg aggccatctt taagcacatg ttctgtgacc
     2581 ccatgtgcgc catcacggag atggaggttg acccctgggt tctgttcggc catccccccg
     2641 ccacccatcc cgacgagctg ctgctccaca aggccaagct ggcctgcggg aacgagttcg
     2701 aggggcgtgt ctgcatagcg ttgcgtgccc tcatctacac cttcaagacg tatcaggtat
     2761 ttgtaccaaa gcccaccgcg ctcgccacat ttgtccgtga ggcgggcgcg ctgcttagac
     2821 gccactcgat ctcgctcctg tccctggagc acaccctgtg tacctatgta tgatcgagtc
     2881 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2941 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3001 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3061 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3121 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3181 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3241 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3301 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3361 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3421 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3481 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3541 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3601 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3781 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3841 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3901 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3961 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
     4021 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
     4081 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
     4141 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
     4201 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
     4261 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
     4321 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
     4381 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
     4441 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4501 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4561 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4621 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4681 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4741 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4801 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4861 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4921 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4981 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5041 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5101 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5161 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5221 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5281 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5341 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5401 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5461 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5521 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5581 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5641 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5701 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5821 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5881 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5941 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6001 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6061 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6121 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6181 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6241 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6301 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6361 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6421 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6481 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6541 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6601 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6661 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6721 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6781 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6841 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6901 tcg
//
