LOCUS       sequence-456524-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral shRNA vector for knockdown of both mouse MKL1 and MKL2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Lee SM, Vasishtha M, Prywes R
  TITLE     Activation and repression of cellular immediate early genes by serum
            response factor cofactors.
  JOURNAL   J Biol Chem. 2010 Jul 16. 285(29):22036-49.
  PUBMED    20466732
REFERENCE   2  (bases 1 to 7084)
  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: "J Biol 
            Chem. 2010 Jul 16. 285(29):22036-49."
COMMENT     Sequence Label: pLKO.1-MKL1/2 shRNA
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(312..911)
                     /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(395..415)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     892..911
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(933..1443)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(996..1014)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    1492..1609
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(1706..1954)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1764..1783)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1934..1954)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(2252..2296)
                     /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    2481..2714
                     /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    3207..3332
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3379..3559
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3560..3786)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(3814..3834)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3814..3832)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3853..3869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3853..3869)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3866..3888)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3877..3893
                     /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(3901..3907,3908..3925,3926..3931))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3946..3967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4084..4101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4255..4843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4335..4354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5014..5805,5806..5874))
                     /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     5637..5656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5875..5979)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6005..6460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6142..6163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6354..6373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6587..6609
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6601..6618
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6602..6618
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6628..6647
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6628..6646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      6667..6802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6842..6963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6853..6872)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6907..6926
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             join(7035..7084,1..184)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
       61 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
      121 cagagagacc cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg aattagccct
      181 tccagtcccc ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt
      241 ggtcttaaag gtaccgatgc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg
      301 tggggcgggc gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
      361 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
      421 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
      481 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
      541 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
      601 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
      661 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
      721 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
      781 tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
      841 gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
      901 tactcggtca tggtaagctc cggtggatcc ccctggggag agaggtcggt gattcggtca
      961 acgagggagc cgactgccga cgtgcgctcc ggaggcttgc agaatgcgga acaccgcgcg
     1021 ggcaggaaca gggcccacac taccgcccca caccccgcct cccgcaccgc cccttcccgg
     1081 ccgctgctct cggcgcgccc tgctgagcag ccgctattgg ccacagccca tcgcggtcgg
     1141 cgcgctgcca ttgctccctg gcgctgtccg tctgcgaggg tactagtgag acgtgcggct
     1201 tccgtttgtc acgtccggca cgccgcgaac cgcaaggaac cttcccgact taggggcgga
     1261 gcaggaagcg tcgccggggg gcccacaagg gtagcggcga agatccgggt gacgctgcga
     1321 acggacgtga agaatgtgcg agacccaggg tcggcgccgc tgcgtttccc ggaaccacgc
     1381 ccagagcagc cgcgtccctg cgcaaaccca gggctgcctt ggaaaaggcg caaccccaac
     1441 cccctcgagc cgcggccaaa gtggatctct gctgtccctg taataaaccc gaaaattttg
     1501 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
     1561 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
     1621 tactgccatt tgtctcgagg tcgagaattc aaaaacatgg agctggtgga gaagaactcg
     1681 agttcttctc caccagctcc atgccggtgt ttcgtccttt ccacaagata tataaagcca
     1741 agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta aaacataatt
     1801 ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt tgtactaata
     1861 tctttgtgtt tacagtcaaa ttaattccaa ttatctctct aacagccttg tatcgtatat
     1921 gcaaatatga aggaatcatg ggaaataggc cctcggtgaa gggggcggcc gctcgaggct
     1981 agtctcgtga tcgataccgt cgagatccgt tcactaatcg aatggatctg tctctgtctc
     2041 tctctccacc ttcttcttct attccttcgg gcctgtcggg tcccctcggg gttgggaggt
     2101 gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata gaaagtacag
     2161 caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc
     2221 acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata
     2281 attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc
     2341 ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt
     2401 ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc
     2461 cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga
     2521 ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct
     2581 caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct
     2641 gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa
     2701 ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct
     2761 tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc
     2821 tccaattgtc cctcatatct cctcctccag gtctgaagat cagcggccgc ttgctgtgcg
     2881 gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     2941 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     3001 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     3061 gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact
     3121 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     3181 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     3241 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     3301 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     3361 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     3421 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     3481 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     3541 ggtctaacca gagagacccg tttatgtatc gagctaggca cttaaataca atatctctgc
     3601 aatgcggcaa ttcagtggtt cgtccaatcc atgtcagacc cgtctgttgc cttcctaata
     3661 aggcacgatc gtaccacctt acttccacca atcggcatgc acggtgcttt ttctctcctt
     3721 gtaaggcatg ttgctaactc atcgttacca tgttgcaaga ctacaagagt attgcataag
     3781 actacattaa gcttgcagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     3841 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3901 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3961 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6001 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6061 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6121 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6181 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6241 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6301 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6361 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6421 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     6481 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6541 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6601 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacg
     6661 tccctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     6721 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     6781 tgggcggagt taggggcggg atgggcggag ttaggggcgg gatagctaga gccagacatg
     6841 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     6901 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     6961 gttcctctca ctctctgata ttcatttctt tgcaagttat aaatactgaa taataagatg
     7021 acatgaacta ctactgctag agattttcca cactgactaa aagggtctga gggatctcta
     7081 gtta
//
