LOCUS       sequence-163173-        7291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing a control sgRNA. NOTE: This plasmid 
            does NOT contain Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7291)
  AUTHORS   Park RJ, Wang T, Koundakjian D, Hultquist JF, Lamothe-Molina P, 
            Monel B, Schumann K, Yu H, Krupzcak KM, Garcia-Beltran W, 
            Piechocka-Trocha A, Krogan NJ, Marson A, Sabatini DM, Lander ES, 
            Hacohen N, Walker BD
  TITLE     A genome-wide CRISPR screen identifies a restricted set of HIV host 
            dependency factors.
  JOURNAL   Nat Genet. 2016 Dec 19. doi: 10.1038/ng.3741.
  PUBMED    27992415
REFERENCE   2  (bases 1 to 7291)
  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: "Nat Genet. 
            2016 Dec 19. doi: 10.1038/ng.3741."
COMMENT     Sequence Label: pLenti-sgCTRL-2
FEATURES             Location/Qualifiers
     source          1..7291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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"
     promoter        1717..1965
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1717..1737
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1888..1907
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1987..2062
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2110..2227
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2296..2507
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2533..3132
                     /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(2533..2552)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3029..3049
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3148..3736
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3201..3221)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3619..3630)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3808..4041
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4119..4240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4156..4175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4210..4229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4280..4415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4435..4454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4436..4454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4464..4481)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4464..4480)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4473..4495)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4622..5077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4709..4728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4919..4940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5103..5207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5208..5276,5277..6068)
                     /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(5426..5445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6239..6827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6728..6747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6981..6998
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7115..7136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7151..7156,7157..7174,7175..7181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7189..7205
                     /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     7194..7216
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7213..7229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7213..7229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7248..7268
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7250..7268
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgatta tacctcttca
     1981 cagctggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     2041 aaaagtggca ccgagtcggt gcttttttga attctagatc ttgagacaaa tggcagtatt
     2101 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2161 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2221 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgagtggct
     2281 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2341 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2401 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2461 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg
     2521 ggatccgcca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     2581 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     2641 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     2701 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     2761 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     2821 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     2881 cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga ccaccagggc
     2941 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     3001 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3061 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     3121 cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     3181 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3241 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3301 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3361 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3421 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3481 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3541 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3601 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3661 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3721 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
     3781 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     3841 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     3901 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     3961 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4021 cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt
     4081 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     4141 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4201 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     4261 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4321 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4381 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     4441 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4501 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4561 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4621 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4681 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4741 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4801 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4861 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4921 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4981 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5041 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     5101 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5161 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5221 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5281 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5341 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5401 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5461 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5521 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5581 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5641 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5701 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5761 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5821 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5881 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5941 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6001 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6061 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6121 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6181 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6841 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6901 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6961 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7021 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     7081 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     7141 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7201 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     7261 actaaaggga acaaaagctg gagctgcaag c
//
