LOCUS       sequence-320642-        6423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses UL32 in mammalian cells with an N-terminal TwinStrep tag 
            and HRV 3C protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  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 6423)
  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: pUE1-TSP-UL32
FEATURES             Location/Qualifiers
     source          1..6423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(814..819,820..843,844..879,880..903)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             910..933
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    2752..3324
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2919..2936)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     regulatory      3326..3335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3606..3625)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3650..3697
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(3784..3801)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3784..3800)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3793..3815)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4009..4028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4128..4150
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4188..4206)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4274..4378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4379..4447,4448..5239)
                     /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     complement(4597..4616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5410..5998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5899..5918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6152..6169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6286..6307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6322..6327,6328..6345,6346..6352)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6360..6376
                     /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)."
     primer_bind     6365..6387
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6384..6400
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6384..6400
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaattctaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gaattcgtac ttcaagaact
      601 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
      661 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
      721 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      781 aacccactgc ttaagcctcc ccgggctcga atgagtgcgt ggagtcatcc tcaattcgag
      841 aaaggtggag gttctggcgg tggatcggga ggttcagcgt ggagccaccc gcagttcgaa
      901 aaatccggac tggaagttct gttccagggg cccatggcaa cttcgccccc cggggtcctg
      961 gcgagcgtcg cggtgtgcga agagtccccc ggcagcagct ggaaggcggg ggcgttcgag
     1021 cgcgcgtatg tggccttcga tcccagcctc ctggctctta acgaggctct ctgcgccgag
     1081 ctcctggcgg ccagtcacgt gataggtgtg ccgcccgtcg gtacgctcga cgaggacgtc
     1141 gccgccgacg tggtcaccgc cccctcaagg gcccgcgggg gggcgggcga cggagggggt
     1201 tcgggcgggc gcggaggacc ccgcaacccg cccccggatc cctgtgggga ggggcttttg
     1261 gacaccgggc ccttttccgc ggcggccatc gacaccttcg cgctggaccg gccctgtctg
     1321 gtctgccgca ccatcgagct gtacaagcaa acctaccgcc tctcgcccca gtgggtggcc
     1381 gattacgcgt ttctctgtgc caaatgcctg ggggcgcccc actgcgccgc tagcatcttc
     1441 gtggccgcgt tcgagtttgt gtacgtcatg gaccgccact ttctgcgcac caagaaggcg
     1501 accctggtcg gctccttcgc acgctttgcc ctcaccataa acgacatcca ccggcacttt
     1561 tttctccact gctgcttccg taccgacggc ggggtgcccg ggcggcatgc ccagaagcaa
     1621 cccaagccct cgccctcccc gggagccgcc aaggtgcagt actccaacta ctcgttcctg
     1681 gcgcagtctg ccacccgggc cctcatcgga accttggcct ccgggggcga ggagggggcg
     1741 gggtcggcgg cgggatccgg cacgcagccg tcgctcacca ccgcgctgat gaactggaag
     1801 gattgcgccc gtctcctgga ctgcacggag ggtcggcgcg ggggcgggga cagctgctgt
     1861 acgcgcgccg ccgctcgcaa cggggaattc gagacggtcg ccggggaccg cgagccggag
     1921 gagtcgccgg atacgtgggc gtacgcggat ctggtcctgc tgctcctcgc cggcaccccc
     1981 gccgtctggg agtcggggcc ccagctgcgc gctgccgcgg aggcccgtcg cgccacggtc
     2041 cgccagtcct gggaagcgca tcgcggggca cgcacgcgcg acgtggctcc gcgctttgcg
     2101 cagtttaccg aacccgatgc ccagcccgac ctcgacctgg gccctctgat ggccaccgtg
     2161 ctgaaacacg gccgggggcg cgggcgcacc ggcggagaat gcctgctgtg taacctgtta
     2221 ctggtgcggg cttactggct ggccctgcgt agactccggg cctccgtggt ccgctactcg
     2281 gaaaacaaca cgagcctctt cgactgtatc gtacccgtcg tcgaccagct cgaggcggac
     2341 cccgagacgc agcccgggga cgggggccgg tttgtgagcc tgctgcgggc cgcggggccc
     2401 gaggccatct ttaagcacat gttctgtgac cccatgtgcg ccatcacgga gatggaggtt
     2461 gacccctggg ttctgttcgg ccatcccccc gccacccatc ccgacgagct gctgctccac
     2521 aaggccaagc tggcctgcgg gaacgagttc gaggggcgtg tctgcatagc gttgcgtgcc
     2581 ctcatctaca ccttcaagac gtatcaggta tttgtaccaa agcccaccgc gctcgccaca
     2641 tttgtccgtg aggcgggcgc gctgcttaga cgccactcga tctcgctcct gtccctggag
     2701 cacaccctgt gtacctatgt atgatgatcg acctgcaggc atgcaagctt gcccctctcc
     2761 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2821 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2881 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     2941 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     3001 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     3061 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3121 ttggatagtt gtggaaagag tcaaatggct cacctcaagc gtattcaaca aggggctgaa
     3181 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     3241 tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     3301 ttcctttgaa aaacacgatg ataatgccac catggagcca gtagatccta gactagagcc
     3361 ctggaagcat ccaggaagtc agcctaaaac tgcttgtacc aattgctatt gtaaaaagtg
     3421 ttgctttcat tgccaagttt gtttcataac aaaagcctta ggcatctcct atggcaggaa
     3481 gaagcggaga cagcgacgaa gagctcatca gaacagtcag actcatcaag cttctctatc
     3541 aaagcaaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat aggggggagg
     3601 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3661 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3721 ggctggcact ctgtcgatac cccaccgaga ccccattggg gagcttgata tcgatagctt
     3781 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3841 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3901 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     3961 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     4021 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4081 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4141 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4201 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4261 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4321 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4381 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4441 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4501 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4561 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4621 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4681 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4741 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4801 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4861 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4921 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4981 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5041 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5101 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5161 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5221 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5281 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5341 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5521 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5641 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5701 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5761 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5821 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5881 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5941 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6001 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6061 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6181 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6241 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6301 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6361 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg aattatcgat
     6421 atc
//
