LOCUS       sequence-383905-        8944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for constructing single casette expression plasmids using 
            the Mammalian Tookit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8944)
  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 8944)
  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 Single Cassette Destination Vec...
FEATURES             Location/Qualifiers
     source          1..8944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             84..717
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    764..889
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1386..1619
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1804..1848
                     /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    2133..2250
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     RBS             2461..2472
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2479..3192
                     /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      3204..3275
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3291..3318
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3351..3367)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3352..3368)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3458..3691
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(3716..3734)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3717..3734)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4024..4043)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4257..4276)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4376..4398
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4436..4454)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      complement(4633..5221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4713..4732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5324..5428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5429..5497,5498..6289)
                     /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(5647..5666)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     6531..6548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6618..6638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6623..6952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6803..6938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          8086..8151
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8281..8301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8726..8860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8763..8782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8817..8836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
       61 atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat
      121 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
      181 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
      241 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
      301 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
      361 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
      421 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
      481 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactggga
      541 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      601 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
      661 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
      721 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
      781 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
      841 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
      901 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
      961 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     1021 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1081 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1141 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1201 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     1261 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1321 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1381 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1441 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1501 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1561 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1621 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1681 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1741 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1801 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1861 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1921 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1981 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2041 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     2101 cggtatcgcc aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     2161 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     2221 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     2281 gtttggccct ctgatgagac gaacgtgaga ccgaaagtga aacgtgattt catgcgtcat
     2341 tttgaacatt ttgtaaatct tatttaataa tgtgtgcggc aattcacatt taatttatga
     2401 atgttttctt aacatcgcgg caactcaaga aacggcaggt tcggatctta gctactagag
     2461 aaagaggaga aatactagat gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt
     2521 ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa
     2581 ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg
     2641 gttccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat
     2701 ccggaccata tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag
     2761 gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt
     2821 gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaggacggc
     2881 aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc
     2941 gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc
     3001 agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg
     3061 ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa
     3121 cgcgatcata tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat
     3181 gaactgtaca aatgaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     3241 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     3301 cgggtgggcc tttctgcgtt tataggtctc agctgcgtct caagcataca cgataccgtc
     3361 gacctcgagg gaattaattc gagctcggta cctttaagac caatgactta caaggcagct
     3421 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3481 cgaagacaag atctgctttt tgcttgtact gggtttctct ggttagacca gatctgagcc
     3541 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3601 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3661 cccttttagt cagtgtggaa aatctctagc agcatctaga attaattccg tgtattctat
     3721 agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg tagccgcgtt
     3781 ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag cagaccctat
     3841 catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc tgagtttctg
     3901 gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc atcgctagcc
     3961 agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     4021 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     4081 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     4141 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
     4201 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg aatggtgcac tctcagtaca
     4261 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4321 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac catctccggg
     4381 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagatg aaagggcctc
     4441 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4501 ggcacttttc ggggaaatgt gccgagcggc cgcacggtta tccacagaat caggggataa
     4561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5281 agggattttg gtcatgacag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5341 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5401 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5461 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5521 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5581 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5641 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5701 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5761 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5821 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5881 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5941 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6001 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6061 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6121 taaatctgga gccggtgagc gtgggtcccg cggtatcatt gcagcactgg ggccagatgg
     6181 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6241 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6301 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6361 ggtgaagatc ctttttgata atcgcggccg ccaatttttt acggttcctg gccttttgct
     6421 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6481 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6541 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6601 ttcattaatg cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     6721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     6781 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6901 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttgg
     6961 acacaagaca ggcttgcgag atatgtttga gaataccact ttatcccgcg tcagggagag
     7021 gcagtgcgta aaaagacgcg gactcatgtg aaatactggt ttttagtgcg ccagatctct
     7081 ataatctcgc gcaacctatt ttcccctcga acacttttta agccgtagat aaacaggctg
     7141 ggacacttca catgagcgaa aaatacatcg tcacctggga catgttgcag atccatgcac
     7201 gtaaactcgc aagccgactg atgccttctg aacaatggaa aggcattatt gccgtaagcc
     7261 gtggcggtct gtaccgggtg cgttactggc gcgtgaactg ggtattcgtc atgtcgatac
     7321 cgtttgtatt tccagctacg atcacgacaa ccagcgcgag cttaaagtgc tgaaacgcgc
     7381 agaaggcgat ggcgaaggct tcatcgttat tgatgacctg gtggataccg gtggtactgc
     7441 ggttgcgatt cgtgaaatgt atccaaaagc gcactttgtc accatcttcg caaaaccggc
     7501 tggtcgtccg ctggttgatg actatgttgt tgatatcccg caagatacct ggattgaaca
     7561 gccgtgggat atgggcgtcg tattcgtccc gccaatctcc ggtcgctaat cttttcaacg
     7621 cctggcactg ccgggcgttg ttctttttaa cttcaggcgg gttacaatag tttccagtaa
     7681 gtattctgga ggctgcatcc atgacacagg caaacctgag cgaaaccctg ttcaaacccc
     7741 gctttaaaca tcctgaaacc tcgacgctag tccgccgctt taatcacggc gcacaaccgc
     7801 ctgtgcagtc ggcccttgat ggtaaaacca tccctcactg gtatcgcatg attaaccgtc
     7861 tgatgtggat ctggcgcggc attgacccac gcgaaatcct cgacgtccag gcacgtattg
     7921 tgatgagcga tgccgaacgt accgacgatg atttatacga tacggtgatt ggctaccgtg
     7981 gcggcaactg gatttatgag tgggccccgg atctttgtga aggaacctta cttctgtggt
     8041 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
     8101 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
     8161 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
     8221 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
     8281 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
     8341 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
     8401 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
     8461 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
     8521 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
     8581 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
     8641 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     8701 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     8761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8821 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     8881 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     8941 tcat
//
