LOCUS       sequence-320571-        6567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal 2x-Strep-tagged BFLF1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6567)
  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 6567)
  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-BFLF1
FEATURES             Location/Qualifiers
     source          1..6567
                     /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(2574..2591)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2580..2804
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2850..3278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2937..2956)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3147..3168
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3287..3307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3292..3621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3472..3607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3534..3553
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3669..3716
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3735..4109
                     /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    4239..4360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4276..4295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4330..4349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4409..4425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4409..4425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4422..4444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4433..4449
                     /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(4457..4463,4464..4481,4482..4487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4502..4523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4640..4657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4811..5399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4891..4910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5570..6361,6362..6430))
                     /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     6193..6212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6431..6535)
                     /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 ccatggagga ctttgtgccc tggaccgtgg acaatctgaa aagccagttc gaggcagtcg
     1141 gccttttgat ggcccactcc tatcttccag ccaacgcgga agaaggcata gcctacccac
     1201 cactcgtcca cacctacgag tccctctctc cagccagcac ctgccgagtt tgtgatctgc
     1261 tcgacaccct ggtcaatcac tcagatgccc ccgtggcctt ctttgaggac tatgccctgc
     1321 tctgctacta ctgtctaaat gccccacgag cctggatctc tagcctcatc acgggcatgg
     1381 actttttaca catcctgata aaatacttcc ccatggccgg ggggttggat tccctcttca
     1441 tgccctccag gattctggcc attgacatcc agcttcactt ttacatttgc cggtgcttcc
     1501 tccccgtgtc ctcttctgac atgattagaa atgccaacct cggctactac aaattggagt
     1561 ttctgaaatc catcctcacc ggccagtccc cggccaactt ctgcttcaag tccatgtggc
     1621 ccagaaccac ccccactttt cttaccctcc ctggacccag aacctgcaaa gattcccagg
     1681 atgtcccagg ggacatcggc aggggccagt acaccgccct gtgctgccac ctgcctacca
     1741 gaaaccgagt tcagcatcca tttctcaggg ccgaaaaggg aggactctct cctgaaatta
     1801 caaccaaggc cgactactgt ggactactat taggtacgtg gcaggggacg gacctgctcg
     1861 gggggccagg ccaccatgct atcggtttaa acgcggaata cagtggcgat gagctggcgg
     1921 agttggccct ggccatcacc agaccggagg ctggcgacca ttcgcagggc ccttgcctcc
     1981 tggcccccat gtttggacta aggcacaaga atgcctcacg gaccatctgc ccactctgcg
     2041 agagcctcgg ggcccacccg gacgccaagg acacccttga ccgcttcaag agcctgattc
     2101 tggactcatt cggcaacaac atcaaaatcc tagacaggat cgtgttcctc atcaagaccc
     2161 aaaacacgct gctggatgtg ccatgcccca ggctaagggc ctggctgcag atgtgcaccc
     2221 cgcaagactt ccacaagcac ctgttctgcg accccctgtg tgccatcaac cacagcatca
     2281 cgaatccgag cgtgctgttt ggtcagattt acccacccag cttccaggcc tttaaggcag
     2341 cgcttgcagc cggtcagaat ctggaacagg gggtctgtga ctcgttaatt acgcttgtgt
     2401 acatatttaa atccacacaa gtggccagag tgggcaaaac aatcctcgtg gatgtcacta
     2461 aggaactgga cgtggtcctg cgcatccacg gccttgacct ggtacagtcc tatcaaactt
     2521 cccaggtcta cgtgtaatcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2581 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2641 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2701 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2761 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2821 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2881 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2941 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3001 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3061 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3121 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3181 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3241 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3301 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3421 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3481 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3541 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3601 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3661 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3721 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     3781 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3841 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3901 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     3961 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     4021 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     4081 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     4141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4381 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4441 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4501 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4561 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4621 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5461 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5521 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5641 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5701 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5761 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5821 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5881 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5941 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6001 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6061 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6541 ttccccgaaa agtgccacct gacgtcg
//
