LOCUS       sequence-320748-        8585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for stable expression of N-terminal 2xStrep-tagged
            BFLF1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8585)
  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 8585)
  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-BFLF1
FEATURES             Location/Qualifiers
     source          1..8585
                     /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             374..397
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             434..457
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1962..2079
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2128..2638
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2557..2575
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2647..3021
                     /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             3093..3326
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3398..3519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3435..3454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3489..3508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3559..3694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3714..3733)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3715..3733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3743..3760)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3743..3759)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3752..3774)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3901..4356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3988..4007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4198..4219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4382..4486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4487..4555,4556..5347)
                     /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(4705..4724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5518..6106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6007..6026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6260..6277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6394..6415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6430..6435,6436..6453,6454..6460)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6468..6484
                     /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     6473..6495
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6492..6508
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6492..6508
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6527..6547
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6529..6547
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6575..6801
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6802..6982
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7029..7154
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7647..7880
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8065..8109
                     /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     8407..8427
                     /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 atgtggagcc acccccagtt cgagaagggc ggcggcagcg gcggcggcag
      421 cggcggcggc agctggagcc acccccagtt cgagaagggg gcggccatgg aggactttgt
      481 gccctggacc gtggacaatc tgaaaagcca gttcgaggca gtcggccttt tgatggccca
      541 ctcctatctt ccagccaacg cggaagaagg catagcctac ccaccactcg tccacaccta
      601 cgagtccctc tctccagcca gcacctgccg agtttgtgat ctgctcgaca ccctggtcaa
      661 tcactcagat gcccccgtgg ccttctttga ggactatgcc ctgctctgct actactgtct
      721 aaatgcccca cgagcctgga tctctagcct catcacgggc atggactttt tacacatcct
      781 gataaaatac ttccccatgg ccggggggtt ggattccctc ttcatgccct ccaggattct
      841 ggccattgac atccagcttc acttttacat ttgccggtgc ttcctccccg tgtcctcttc
      901 tgacatgatt agaaatgcca acctcggcta ctacaaattg gagtttctga aatccatcct
      961 caccggccag tccccggcca acttctgctt caagtccatg tggcccagaa ccacccccac
     1021 ttttcttacc ctccctggac ccagaacctg caaagattcc caggatgtcc caggggacat
     1081 cggcaggggc cagtacaccg ccctgtgctg ccacctgcct accagaaacc gagttcagca
     1141 tccatttctc agggccgaaa agggaggact ctctcctgaa attacaacca aggccgacta
     1201 ctgtggacta ctattaggta cgtggcaggg gacggacctg ctcggggggc caggccacca
     1261 tgctatcggt ttaaacgcgg aatacagtgg cgatgagctg gcggagttgg ccctggccat
     1321 caccagaccg gaggctggcg accattcgca gggcccttgc ctcctggccc ccatgtttgg
     1381 actaaggcac aagaatgcct cacggaccat ctgcccactc tgcgagagcc tcggggccca
     1441 cccggacgcc aaggacaccc ttgaccgctt caagagcctg attctggact cattcggcaa
     1501 caacatcaaa atcctagaca ggatcgtgtt cctcatcaag acccaaaaca cgctgctgga
     1561 tgtgccatgc cccaggctaa gggcctggct gcagatgtgc accccgcaag acttccacaa
     1621 gcacctgttc tgcgaccccc tgtgtgccat caaccacagc atcacgaatc cgagcgtgct
     1681 gtttggtcag atttacccac ccagcttcca ggcctttaag gcagcgcttg cagccggtca
     1741 gaatctggaa cagggggtct gtgactcgtt aattacgctt gtgtacatat ttaaatccac
     1801 acaagtggcc agagtgggca aaacaatcct cgtggatgtc actaaggaac tggacgtggt
     1861 cctgcgcatc cacggccttg acctggtaca gtcctatcaa acttcccagg tctacgtgta
     1921 aaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     1981 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     2041 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     2101 agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc aaggcagccc
     2161 tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca gcggcgccga
     2221 ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct tcgccgctac
     2281 ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag gttccttgcg
     2341 gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta ccctcgcaga
     2401 cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg
     2461 ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg
     2521 tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc
     2581 ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg
     2641 ggatccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     2701 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     2761 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     2821 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     2881 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     2941 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     3001 gtggccgagg agcaggactg aggtaccttt aagaccaatg acttacaagg cagctgtaga
     3061 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     3121 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     3181 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     3241 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     3301 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     3361 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     3421 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     3541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3661 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     3721 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3781 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3841 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     3901 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3961 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4021 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4081 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4141 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4201 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4261 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4321 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4381 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4441 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4501 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4561 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4621 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4681 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4741 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4801 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4861 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4921 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4981 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5041 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5101 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5161 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5221 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5281 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5341 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5401 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5461 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5521 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5581 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5641 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5701 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5761 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5821 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5881 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5941 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6001 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6061 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6241 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6301 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6361 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6421 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6481 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     6541 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     6601 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     6661 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     6721 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     6781 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     6841 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6901 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6961 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     7021 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     7081 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     7141 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     7201 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7261 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7321 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7381 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7441 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7501 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     7561 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     7621 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     7681 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     7741 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     7801 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     7861 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     7921 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     7981 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     8041 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     8101 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     8161 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     8221 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     8281 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     8341 cgattagtga acggatctcg acggtatcga tcacgagact agcctcgagc ggccgccccc
     8401 ttcaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     8461 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     8521 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     8581 tatca
//
