LOCUS       sequence-139509-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hairpin targeting MEK1 and MEK2 (murine).  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Jensen KJ, Moyer CB, Janes KA
  TITLE     Network Architecture Predisposes an Enzyme to Either Pharmacologic 
            or Genetic Targeting.
  JOURNAL   Cell Syst. 2016 Feb 24;2(2):112-121.
  PUBMED    26942229
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"; journalName: "Cell 
            Syst."; date: "2016-02-24"; volume: "2(2)"; pages: "112-121"
COMMENT     Sequence Label: shMEK1/2 puro
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     425..443
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             528..1127
                     /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(528..547)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1024..1044
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1255..1488
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1560..1681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1597..1616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1651..1670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1721..1856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1876..1895)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1877..1895)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1905..1922)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1905..1921)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1914..1936)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2063..2518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2150..2169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2360..2381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2544..2648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2649..2717,2718..3509)
                     /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(2867..2886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3680..4268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4169..4188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4422..4439
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4556..4577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4592..4597,4598..4615,4616..4622)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4630..4646
                     /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     4635..4657
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4654..4670
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4654..4670
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4689..4709
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4691..4709
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4737..4963
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4964..5144
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5191..5316
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5809..6042
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6227..6271
                     /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        6569..6817
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6569..6589
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6740..6759
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6914..7031
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(7080..7083,1..506)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg
      421 cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg
      481 accgaatcac cgacctctct ccccaggggg atccaccgga gcttaccatg accgagtaca
      541 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
      601 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
      661 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
      721 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
      781 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
      841 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
      901 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
      961 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     1021 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     1081 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     1141 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcggtacct ttaagaccaa
     1201 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     1261 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     1321 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     1381 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     1441 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     1501 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     1561 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1621 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1681 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     1741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1801 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     1861 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     1921 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     1981 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2041 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2101 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2161 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2221 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2281 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2341 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2401 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2461 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     2521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2641 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2761 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2821 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2881 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2941 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3061 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3181 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3241 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3481 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3541 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3601 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3661 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4321 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4381 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4441 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4501 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4561 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4621 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4681 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
     4741 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
     4801 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
     4861 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
     4921 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
     4981 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5041 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5101 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     5161 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     5221 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     5281 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     5341 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     5401 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     5461 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     5521 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     5581 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     5641 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     5701 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     5761 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     5821 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     5881 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     5941 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     6001 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     6061 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     6121 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     6181 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     6241 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     6301 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     6361 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     6421 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     6481 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gatcacgaga
     6541 ctagcctcga gcggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt
     6601 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     6661 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     6721 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     6781 cttggcttta tatatcttgt ggaaaggacg aaacaccggc cagatcatcc gggagctgca
     6841 ctgcagtgca gctcccggat gatctggttt ttgaattctc gacctcgaga caaatggcag
     6901 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     6961 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     7021 ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggccg cggctcgagg
     7081 ggg
//
