LOCUS       sequence-350784-        7050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control shRNA vector containing non-hairpin insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7050)
  AUTHORS   Moffat J, Grueneberg DA, Yang X, Kim SY, Kloepfer AM, Hinkle G, 
            Piqani B, Eisenhaure TM, Luo B, Grenier JK, Carpenter AE, Foo SY, 
            Stewart SA, Stockwell BR, Hacohen N, Hahn WC, Lander ES, Sabatini 
            DM, Root DE
  TITLE     A lentiviral RNAi library for human and mouse genes applied to an 
            arrayed viral high-content screen.
  JOURNAL   Cell. 2006 Mar 24. 124(6):1283-98.
  PUBMED    16564017
REFERENCE   2  (bases 1 to 7050)
  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: "Cell. 2006 
            Mar 24. 124(6):1283-98."
COMMENT     Sequence Label: pLKO.1 - TRC control
FEATURES             Location/Qualifiers
     source          1..7050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(955..1746,1747..1815))
                     /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     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1920)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1946..2401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2083..2104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2295..2314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2528..2550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2542..2559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2543..2559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2569..2588
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2569..2587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2608..2743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2783..2904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2794..2813)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2848..2867
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2976..3209
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(3337..3936)
                     /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(3420..3440)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3917..3936
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3958..4468)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4021..4039)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    4517..4634
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(4697..4945)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4755..4774)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4925..4945)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(5243..5287)
                     /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    5472..5705
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6198..6323
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6370..6550
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6551..6777)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(6805..6825)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6805..6823)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6844..6860)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6844..6860)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6857..6879)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6868..6884
                     /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)."
     promoter        complement(join(6892..6898,6899..6916,6917..6922))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6937..6958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1921 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     1981 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2041 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2101 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2161 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2221 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2281 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2341 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2401 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     2461 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     2521 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     2581 ctatagggcg aattgggtac gtccctaggc ctccaaaaaa gcctcctcac tacttctgga
     2641 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     2701 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     2761 ggatagctag agccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2821 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2881 attataagct gcaataaaca agttcctctc actctctgat attcatttct ttgcaagtta
     2941 taaatactga ataataagat gacatgaact actactgcta gagattttcc acactgacta
     3001 aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac acactacttg
     3061 aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag
     3121 ctcccaggct cagatctggt ctaaccagag agacccagta caagcaaaaa gcagatcttg
     3181 tcttcgttgg gagtgaatta gcccttccag tccccccttt tcttttaaaa agtggctaag
     3241 atctacagct gccttgtaag tcattggtct taaaggtacc gatgcatggg gtcgtgcgct
     3301 cctttcggtc gggcgctgcg ggtcgtgggg cgggcgtcag gcaccgggct tgcgggtcat
     3361 gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg
     3421 ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg
     3481 ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc
     3541 gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag
     3601 gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg
     3661 gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac
     3721 cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa
     3781 gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt
     3841 ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg
     3901 ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggta agctccggtg gatccccctg
     3961 gggagagagg tcggtgattc ggtcaacgag ggagccgact gccgacgtgc gctccggagg
     4021 cttgcagaat gcggaacacc gcgcgggcag gaacagggcc cacactaccg ccccacaccc
     4081 cgcctcccgc accgcccctt cccggccgct gctctcggcg cgccctgctg agcagccgct
     4141 attggccaca gcccatcgcg gtcggcgcgc tgccattgct ccctggcgct gtccgtctgc
     4201 gagggtacta gtgagacgtg cggcttccgt ttgtcacgtc cggcacgccg cgaaccgcaa
     4261 ggaaccttcc cgacttaggg gcggagcagg aagcgtcgcc ggggggccca caagggtagc
     4321 ggcgaagatc cgggtgacgc tgcgaacgga cgtgaagaat gtgcgagacc cagggtcggc
     4381 gccgctgcgt ttcccggaac cacgcccaga gcagccgcgt ccctgcgcaa acccagggct
     4441 gccttggaaa aggcgcaacc ccaaccccct cgagccgcgg ccaaagtgga tctctgctgt
     4501 ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt
     4561 gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc
     4621 cttttctttt aaaattgtgg atgaatactg ccatttgtct cgaggtcgag aattcacgcg
     4681 tgcatacctg cggaccggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga
     4741 aatactttca agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg
     4801 caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt
     4861 ttacagtcaa attaattcca attatctctc taacagcctt gtatcgtata tgcaaatatg
     4921 aaggaatcat gggaaatagg ccctcggtga agggggcggc cgctcgaggc tagtctcgtg
     4981 atcgataccg tcgagatccg ttcactaatc gaatggatct gtctctgtct ctctctccac
     5041 cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa
     5101 acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta
     5161 ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat
     5221 ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt
     5281 cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat
     5341 tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt
     5401 tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac
     5461 cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct
     5521 ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc
     5581 tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca
     5641 gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag
     5701 ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg
     5761 ctactcctaa tggttcaatt tttactactt tatatttata taattcactt ctccaattgt
     5821 ccctcatatc tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta
     5881 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     5941 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     6001 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     6061 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     6121 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     6181 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     6241 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     6301 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     6361 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     6421 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     6481 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     6541 agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg caatgcggca
     6601 attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat aaggcacgat
     6661 cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct tgtaaggcat
     6721 gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa gactacatta
     6781 agcttgcagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     6841 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6961 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7021 aatcggccaa cgcgcgggga gaggcggttt
//
