LOCUS       sequence-320802-        8534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for stable expression of N-terminal 2xStrep-tagged
            ORF68 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8534)
  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 8534)
  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-ORF68
FEATURES             Location/Qualifiers
     source          1..8534
                     /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"
     regulatory      741..750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1911..2028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2077..2587
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2506..2524
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2596..2970
                     /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             3042..3275
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3347..3468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3384..3403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3438..3457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3508..3643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3663..3682)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3664..3682)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3692..3709)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3692..3708)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3701..3723)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3850..4305
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3937..3956)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4147..4168
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4331..4435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4436..4504,4505..5296)
                     /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(4654..4673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5467..6055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5956..5975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6209..6226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6343..6364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6379..6384,6385..6402,6403..6409)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6417..6433
                     /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     6422..6444
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6441..6457
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6441..6457
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6476..6496
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6478..6496
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6524..6750
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6751..6931
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6978..7103
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7596..7829
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8014..8058
                     /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     8356..8376
                     /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 gcggccatgt ttgttccctg
      481 gcaactcggt acaattaccc gtcaccgaga tgagctccaa aaactactgg cagcctccct
      541 gctcccggag cacccggagg agagcctcgg taaccccata atgacacaga ttcaccagtc
      601 gctccaacca tcttccccct gcagggtctg tcagctccta ttttctctgg tccgcgattc
      661 gtccaccccc atgggtttct tcgaggacta tgcctgcctc tgcttcttct gtctatacgc
      721 cccacactgc tggacctcga ccatggcggc agcggcagac ctgtgcgaga tcatgcatct
      781 gcactttcca gaagaggagg cgacatacgg gctattcgga ccgggtcgcc ttatgggtat
      841 cgacttgcag ctgcacttct ttgttcaaaa gtgctttaag accaccgccg ccgaaaaaat
      901 actgggaata tccaacctgc aatttttaaa atcagaattc atccggggca tgctcacagg
      961 caccatcacc tgcaacttct gcttcaaaac gtcctggccc aggacagaca aggaggaggc
     1021 caccggcccc accccatgct gccagattac agacaccacc accgcacccg cgagcggcat
     1081 accggaacta gcccgggcca cattctgcgg cgcaagtcgc cccacaaagc ccagcctact
     1141 tcccgcgcta atagatatct ggtccacgag ctcagagctc cttgacgagc cgcgccctcg
     1201 actgatcgca agcgacatga gtgaactcaa atccgtggtc gcatcccacg atccgttctt
     1261 ctctcccccg cttcaggcag acacctccca gggtccatgt ctaatgcacc caaccctggg
     1321 gctacgatac aaaaacggga ctgcatccgt ctgcctcctc tgcgagtgcc ttgcggcaca
     1381 cccagaggca cccaaggcgc tgcagaccct tcagtgcgag gtaatgggcc atatagaaaa
     1441 caacgtaaag ctggtagaca gaattgcctt tgtgttggac aacccattcg ccatgccata
     1501 tgtctcagat ccgctactta gagagctgat ccggggctgt accccacagg aaattcacaa
     1561 gcacctgttc tgcgacccgc tgtgcgccct caatgctaag gtggtgtcag aggacgtact
     1621 attccgcctg cccagggagc aggagtataa aaagctcagg gcatccgcgg ccgccggaca
     1681 gctcctcgat gccaacaccc tgttcgactg cgaggtcgtg cagactttgg tctttctctt
     1741 taagggtctc caaaacgcca gggtggggaa aaccacctca ctagacatta ttcgggagct
     1801 aaccgcacaa ctaaaaagac accgcctaga cctggcccac ccctcacaga cgtcacactt
     1861 gtacgcttaa aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     1921 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     1981 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     2041 agggacagca gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca
     2101 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     2161 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     2221 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     2281 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     2341 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     2401 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     2461 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     2521 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     2581 tccccagggg gatccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     2641 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     2701 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     2761 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     2821 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     2881 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     2941 gtgcacttcg tggccgagga gcaggactga ggtaccttta agaccaatga cttacaaggc
     3001 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     3061 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     3121 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3181 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3241 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     3301 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     3361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     3481 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3541 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3601 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     3661 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     3721 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3781 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3841 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3901 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3961 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4021 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4081 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4141 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4201 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4261 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4321 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4381 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4441 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4501 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4561 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4621 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4681 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4741 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4801 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4861 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4921 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4981 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5041 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5101 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5161 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5221 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5281 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5581 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6181 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6241 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6301 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     6361 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6421 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     6481 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     6541 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     6601 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     6661 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     6721 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     6781 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6841 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6901 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     6961 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     7021 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     7081 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     7141 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     7201 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     7261 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     7321 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     7381 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     7441 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     7501 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     7561 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     7621 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     7681 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     7741 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     7801 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     7861 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     7921 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     7981 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     8041 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     8101 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     8161 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     8221 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     8281 agatccattc gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg
     8341 gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac
     8401 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     8461 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     8521 aaaatggact atca
//
