LOCUS       sequence-320798-        8445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for stable expression of N-terminal 2xStrep-tagged
            mu68 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8445)
  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 8445)
  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: pLJM1-zeo-2xStrep-mu68
FEATURES             Location/Qualifiers
     source          1..8445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             375..398
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             435..458
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1988..2498
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2417..2435
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2507..2881
                     /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"
     LTR             2953..3186
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3258..3379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3295..3314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3349..3368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3419..3554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3574..3593)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3575..3593)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3603..3620)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3603..3619)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3612..3634)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3761..4216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3848..3867)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4058..4079
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4242..4346
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4347..4415,4416..5207)
                     /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(4565..4584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5378..5966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5867..5886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6120..6137
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6254..6275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6290..6295,6296..6313,6314..6320)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6328..6344
                     /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     6333..6355
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6352..6368
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6352..6368
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6387..6407
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6389..6407
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6435..6661
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6662..6842
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6889..7014
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7507..7740
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7925..7969
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     8267..8287
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg gatgtggagc cacccccagt tcgagaaggg cggcggcagc ggcggcggca
      421 gcggcggcgg cagctggagc cacccccagt tcgagaaggg ggcggccatg tacgtaccat
      481 ggtctctaga gacagtcacc tcccacgatt caatcatgca gatgttactt tctaaatcct
      541 acctacctgc cacaccagaa acaggctttc atgatcccac actcactcat atctcaagca
      601 cgttgttgcc cacttccaag tgcaagatat gtaacatgct tttcgaagct gtgctggaac
      661 acaatccccc tgattctttt tttagtgagt atgcgaccct ctgcttctac gtgctgcatg
      721 cccccgtgac ctggtccgca acctacataa caatctgtga tatgttgcaa atcttctcta
      781 acgcgggact aaaccatctc attactgaga aaataaactc tagcaagatc atgggcatag
      841 acttttacct tcatttcttt gccaggcaat gcttcaggca caccaccccc gatcaggtgg
      901 ctcaactgga aaacctgcac caggtccgcc tgagtttgtt atctaatacc atctgtaatg
      961 tccagtctgc tgctcacata caaggtatct ggtccagctc tctggaaaca gcccacaaga
     1021 agggggccca gagtccagat ccccccactg gctgccctgt caaggctgtg ccccaccgtg
     1081 agttcttttc aaacaccaac aggccaagtc ttctcaagat attcagcgag gcctggaagc
     1141 ggtccctgtt actcgacccc gattactccg agccagacac acctgtagag acggtggata
     1201 cctcccaagg accgtgccta ctctcgtttc ccttctccct caaatgtagg ggcaacacca
     1261 gctccgtgtg cgtgttgtgc gaatgtctgg cgtgtcaccc cgaggcggcc aggatcctcc
     1321 aagagatcaa atccaccata ctcacctgcc tggaaaataa cacctctttg ctagacagaa
     1381 ccgcctacac cctgatgatg tacaccacct ctaagctcat agatgatcca gacctggctg
     1441 aaatcatcac cagtcgggga cccaatgtac tacataagca ccttttctgt gatccattct
     1501 gtgccataaa cagtctgaaa actgaccctc tggtgctatt taaatatccc agcggaccca
     1561 aatttcaaat ttttacaaaa gccctcgctt ctggaaaata tttggcggag aacgcactat
     1621 ttgaccagcc agaaacagag ttgttagtgt tggcgctgaa ggcctgtcag ttggcacagt
     1681 ttccaaaaaa atttgaagca gaagtgttaa aaggaatttc caggttggga actacgctca
     1741 ccactgtcag tcctgtgcat ttcttttctg tttacaccta aaattctcga cctcgagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac tttggccgcg
     1981 gctcgagggg gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg
     2041 gctgctctgg gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct
     2101 tcacgtccgt tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga
     2161 cgcttcctgc tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt
     2221 gacaaacgga agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa
     2281 tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg
     2341 agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc
     2401 tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg
     2461 gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccatgg ccaagttgac
     2521 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     2581 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     2641 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     2701 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     2761 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     2821 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     2881 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     2941 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     3001 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     3061 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     3121 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     3181 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     3241 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3301 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3361 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     3421 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3481 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3541 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     3601 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3661 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3721 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3781 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3841 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3901 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3961 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4021 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4081 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4141 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4201 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4261 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4321 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4381 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4441 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4501 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4561 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4621 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4681 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4741 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4801 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4861 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4921 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4981 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5041 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5101 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5161 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5221 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5281 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5341 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5401 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5461 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5521 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5581 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5641 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5701 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5761 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5821 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5881 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5941 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6001 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6061 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6121 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6181 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6241 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6301 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6361 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     6421 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     6481 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     6541 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     6601 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     6661 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6721 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6781 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6841 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     6901 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     6961 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     7021 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     7081 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     7141 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     7201 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     7261 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     7321 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     7381 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     7441 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     7501 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     7561 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     7621 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     7681 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     7741 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     7801 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     7861 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     7921 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     7981 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     8041 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     8101 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     8161 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
     8221 acggtatcga tcacgagact agcctcgagc ggccgccccc ttcaccgagg gcctatttcc
     8281 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa
     8341 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     8401 tgggtagttt gcagttttaa aattatgttt taaaatggac tatca
//
