LOCUS       sequence-141827-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shETV4 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Wang B, Krall EB, Aguirre AJ, Kim M, Widlund HR, Doshi MB, Sicinska 
            E, Sulahian R, Goodale A, Cowley GS, Piccioni F, Doench JG, Root DE,
            Hahn WC
  TITLE     ATXN1L, CIC, and ETS Transcription Factors Modulate Sensitivity to 
            MAPK Pathway Inhibition.
  JOURNAL   Cell Rep. 2017 Feb 7;18(6):1543-1557. doi: 
            10.1016/j.celrep.2017.01.031.
  PUBMED    28178529
REFERENCE   2  (bases 1 to 7083)
  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"; doi: 
            "10.1016/j.celrep.2017.01"; journalName: "Cell Rep"; date: 
            "2017-02-7- 7"; volume: "18"; issue: "6"; pages: "1543-1557"
COMMENT     Sequence Label: pLKO.1 shETV4-A
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        207..717
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     636..654
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             739..1338
                     /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(739..758)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1235..1255
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1466..1699
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1771..1892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1808..1827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1862..1881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1932..2067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2087..2106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2088..2106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2116..2133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2116..2132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2125..2147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2274..2729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2361..2380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2571..2592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2755..2859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2860..2928,2929..3720)
                     /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(3078..3097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3891..4479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4380..4399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4633..4650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4767..4788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4803..4808,4809..4826,4827..4833)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4841..4857
                     /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     4846..4868
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4865..4881
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4865..4881
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4900..4920
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4902..4920
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4948..5174
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5175..5355
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5402..5527
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6020..6253
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6438..6482
                     /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        6780..7028
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6780..6800
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6951..6970
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct
      241 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
      301 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
      361 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
      421 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
      481 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
      541 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
      601 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
      661 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
      721 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
      781 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
      841 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
      901 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
      961 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     1021 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     1081 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     1141 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     1201 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     1261 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     1321 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     1381 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     1441 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     1501 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     1561 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     1621 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     1681 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     1741 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     1801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     1921 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1981 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2041 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     2101 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     2281 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2341 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2401 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2461 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2521 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2581 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2641 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2701 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     2761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3061 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3181 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3241 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3301 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3361 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3421 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3481 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3541 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3601 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3661 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3721 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3781 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3841 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3901 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3961 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4021 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4081 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4141 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4201 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4261 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4321 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4381 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4441 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4501 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4561 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4621 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4681 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4741 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4801 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4861 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     4921 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     4981 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     5041 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     5101 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     5161 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5221 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5281 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5341 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     5401 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     5461 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     5521 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     5581 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     5641 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     5701 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     5761 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     5821 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     5881 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     5941 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     6001 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     6061 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     6121 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     6181 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     6241 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     6301 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     6361 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     6421 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     6481 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     6541 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     6601 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     6661 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     6721 tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg
     6781 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     6841 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     6901 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     6961 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     7021 gaaacaccgg cccaacaaat gcccatttca tctcgagatg aaatgggcat ttgttgggtt
     7081 ttt
//
