LOCUS       sequence-384572-        8849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for constructing multi casette expression plasmids using 
            the Mammalian Tookit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8849)
  AUTHORS   Weinberg ZY, Hilburger CE, Kim M, Cao L, Khalid M, Elmes S, Diwanji 
            D, Hernandez E, Lopez J, Schaefer K, Smith AM, Zhou F, QCRG 
            Structural Biology Consortium, Kumar GR, Ott M, Baker D, El-Samad H
  TITLE     Sentinel cells enable genetic detection of SARS-CoV-2 Spike protein
  JOURNAL   bioRxiv 2021.04.20.440678
REFERENCE   2  (bases 1 to 8849)
  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: "bioRxiv 
            2021.04.20.440678"
COMMENT     Sequence Label: MTK Lentivirus Multi Cassette Destination Vect...
FEATURES             Location/Qualifiers
     source          1..8849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             92..725
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    772..897
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1394..1627
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1812..1856
                     /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"
     misc_feature    2141..2258
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     RBS             2469..2480
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2487..3200
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3212..3283
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3299..3326
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3359..3375)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3360..3376)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3466..3699
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(3724..3742)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3725..3742)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4032..4051)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4265..4284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4384..4406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4444..4462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      complement(4641..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4721..4740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5443..5462)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5454..6269
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5454..6269
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5527..5546)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6444..6461
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6531..6551)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6536..6865
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6716..6851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          7999..8064
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8194..8214
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8639..8773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8676..8695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8730..8749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta
       61 tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga
      121 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
      181 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
      241 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
      301 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
      361 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
      421 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
      481 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
      541 tactgggact ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
      601 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
      661 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
      721 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
      781 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
      841 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
      901 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
      961 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     1021 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     1081 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     1141 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     1201 gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg
     1261 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1321 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1381 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1441 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1501 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1561 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1621 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     1681 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     1741 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     1801 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     1861 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     1921 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     1981 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     2041 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     2101 gatctcgacg gtatcgccaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2161 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2221 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2281 gagatccagt ttggccctct gatgagacga acgtgagacc gaaagtgaaa cgtgatttca
     2341 tgcgtcattt tgaacatttt gtaaatctta tttaataatg tgtgcggcaa ttcacattta
     2401 atttatgaat gttttcttaa catcgcggca actcaagaaa cggcaggttc ggatcttagc
     2461 tactagagaa agaggagaaa tactagatgc gtaaaggcga agagctgttc actggtgtcg
     2521 tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
     2581 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
     2641 aactgccggt tccttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
     2701 ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
     2761 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
     2821 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     2881 aggacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     2941 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     3001 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     3061 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     3121 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
     3181 gtatggatga actgtacaaa tgaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3241 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     3301 ctcaccttcg ggtgggcctt tctgcgttta taggtctcag ctgcgtctca agcatacacg
     3361 ataccgtcga cctcgaggga attaattcga gctcggtacc tttaagacca atgacttaca
     3421 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     3481 actcccaacg aagacaagat ctgctttttg cttgtactgg gtttctctgg ttagaccaga
     3541 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     3601 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     3661 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catctagaat taattccgtg
     3721 tattctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta
     3781 gccgcgttct aacgacaata tgtacaagcc taattgtgta gcatctggct tactgaagca
     3841 gaccctatca tctctctcgt aaactgccgt cagagtcggt ttggttggac gaaccttctg
     3901 agtttctggt aacgccgtcc cgcacccgga aatggtcagc gaaccaatca gcagggtcat
     3961 cgctagccag atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     4021 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     4081 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     4141 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     4201 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc
     4261 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4321 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgacca
     4381 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagatgaa
     4441 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4501 cgtcaggtgg cacttttcgg ggaaatgtgc cgagcggccg cacggttatc cacagaatca
     4561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5041 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5281 tcacgttaag ggattttggt catgactgat ccttcaactc agcaaaagtt cgatttattc
     5341 aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     5401 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     5461 atattcaacg ggaaacgtct tgctccaggc cgcgattaaa ttccaacatg gatgctgatt
     5521 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     5581 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     5641 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     5701 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     5761 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     5821 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     5881 gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     5941 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     6001 ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     6061 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     6121 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     6181 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     6241 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacacgcgg
     6301 ccgccaattt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6361 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6421 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6481 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg
     6541 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6601 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6661 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6721 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6781 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6841 ttttggaggc ctaggctttt gcaaaaagct tggacacaag acaggcttgc gagatatgtt
     6901 tgagaatacc actttatccc gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat
     6961 gtgaaatact ggtttttagt gcgccagatc tctataatct cgcgcaacct attttcccct
     7021 cgaacacttt ttaagccgta gataaacagg ctgggacact tcacatgagc gaaaaataca
     7081 tcgtcacctg ggacatgttg cagatccatg cacgtaaact cgcaagccga ctgatgcctt
     7141 ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact
     7201 ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt atttccagct acgatcacga
     7261 caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt
     7321 tattgatgac ctggtggata ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa
     7381 agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt
     7441 tgttgatatc ccgcaagata cctggattga acagccgtgg gatatgggcg tcgtattcgt
     7501 cccgccaatc tccggtcgct aatcttttca acgcctggca ctgccgggcg ttgttctttt
     7561 taacttcagg cgggttacaa tagtttccag taagtattct ggaggctgca tccatgacac
     7621 aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa acatcctgaa acctcgacgc
     7681 tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa
     7741 ccatccctca ctggtatcgc atgattaacc gtctgatgtg gatctggcgc ggcattgacc
     7801 cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg
     7861 atgatttata cgatacggtg attggctacc gtggcggcaa ctggatttat gagtgggccc
     7921 cggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca
     7981 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg
     8041 attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg
     8101 tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat
     8161 gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac
     8221 cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga
     8281 actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa
     8341 attatggaaa aatattctgt aacctttata agtaggcata acagttataa tcataacata
     8401 ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa
     8461 ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt
     8521 gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     8581 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     8641 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8701 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8761 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     8821 ataccctatt accactgcca attacctgt
//
