LOCUS       Exported                8392 bp ds-DNA     circular SYN 17-SEP-2021
DEFINITION  Lentiviral expression of GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pSin-GFP-Fg
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8392)
  AUTHORS   Yue T, Zhan X, Zhang D, Jain R, Wang KW, Choi JH, Misawa T, Su L, 
            Quan J, Hildebrand S, Xu D, Li X, Turer E, Sun L, Moresco EMY, 
            Beutler B
  TITLE     SLFN2 protection of tRNAs from stress-induced cleavage is essential 
            for T cell-mediated immunity.
  JOURNAL   Science. 2021 May 14;372(6543). pii: 372/6543/eaba4220. doi: 
            10.1126/science.aba4220.
  PUBMED    33986151
REFERENCE   2  (bases 1 to 8392)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8392
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     182..201
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             394..993
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(394..413)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     890..910
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1196..1429
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1580..1597)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1586..1810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      1853..1862
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        1906..2010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2011..2871
                     /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(2229..2248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3042..3630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3531..3550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             3789..4422
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    4469..4594
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5087..5320
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5505..5549
                     /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    5836..5953
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6082..7260
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6313..7251
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7208..7228
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7283..7999
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             7283..7999
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7328..7349)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7589..7608)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7936..7957
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8006..8071
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    8180..361
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8347..8364)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
       61 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
      121 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
      181 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
      241 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
      301 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
      361 agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc cacggtgcgc
      421 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
      481 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
      541 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
      601 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
      661 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
      721 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
      781 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
      841 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
      901 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
      961 acctggtgca tgacccgcaa gcccggtgcc tgataatagg cggccgctcg agacctagaa
     1021 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
     1081 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
     1141 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     1201 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     1261 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     1321 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     1381 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     1441 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     1501 ccttgacatt ataatagatt tagcaggaat tgaactagga gtggagcaca caggcaaagt
     1561 tctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1621 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1681 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1741 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1801 ggctctatgg cttctgaggc ggaaagaacc agctgggggc gcgcccctcg aggccgccat
     1861 ggtcatagct gtttgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1921 tgtttatttt tctaaataca ttcaaatatg tatccgctca cgagacaata accctgataa
     1981 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2041 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2101 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2161 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2221 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2281 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2341 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2401 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2461 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2521 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2581 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2641 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2701 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2761 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2821 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2881 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2941 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3001 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3061 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3181 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3661 tggccttttg cacggaccgc cactgccaat tacctgtggt ttcatttact ctaaacctgt
     3721 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
     3781 ttagtagttg gaagggctaa ttcactccca aagaagacaa gatatccttg atctgtggat
     3841 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     3901 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     3961 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     4021 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     4081 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     4141 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     4201 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     4261 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     4321 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     4381 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     4441 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     4501 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     4561 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     4621 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     4681 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     4741 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     4801 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     4861 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     4921 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     4981 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     5041 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     5101 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     5161 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     5221 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     5281 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     5341 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     5401 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     5461 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     5521 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     5581 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     5641 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     5701 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     5761 gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgc cacaaatggc
     5821 agtattcatc cacaatttta aaagaaaggg ggggattggg gggtacagtg caggggaaag
     5881 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     5941 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttgga tcgataagct
     6001 ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag
     6061 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     6121 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
     6181 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     6241 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     6301 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
     6361 ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag
     6421 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
     6481 cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac
     6541 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
     6601 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
     6661 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
     6721 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     6781 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     6841 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     6901 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     6961 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     7021 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     7081 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg
     7141 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     7201 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga
     7261 ggaattgatc cttcgaacta gtatggtgag caagggcgag gagctgttca ccggggtggt
     7321 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     7381 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     7441 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     7501 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     7561 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     7621 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     7681 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     7741 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     7801 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     7861 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     7921 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     7981 catggacgag ctgtacaagc atatggacta caaagaccat gacggtgatt ataaagatca
     8041 tgacatcgac tacaaggatg acgatgacaa ggctagctag tgagaattcg aagcttgagc
     8101 tcgagatctg agtccggtag cgctagttcg aaggatccgc atgcatctag ggcggccaat
     8161 tccgcccctc tccccccccc ccctctccct cccccccccc taacgttact ggccgaagcc
     8221 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     8281 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     8341 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gc
//
