LOCUS       sequence-474984-        4406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1-shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4406)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4406)
  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"
COMMENT     Sequence Label: pSUPERpuro-HsNot1_Q
FEATURES             Location/Qualifiers
     source          1..4406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     674..690
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(753..1352)
                     /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(836..856)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1333..1352
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1377..1876)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1425..1444)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1818..1841
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        1887..2101
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2022..2041
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(2173..2189)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2174..2190)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2218..2236)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2257..2273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2257..2273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2270..2292)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2281..2297
                     /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(2305..2311,2312..2329,2330..2335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2350..2371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2488..2505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2659..3247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2739..2758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3418..4209,4210..4278))
                     /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     4041..4060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4279..4383)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccgggctg catggggtcg tgcgctcctt
      721 tcggtcgggc gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac
      781 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
      841 tagaagggga ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg
      901 gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
      961 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     1021 ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     1081 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     1141 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     1201 tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg
     1261 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg
     1321 gcgaggcgca ccgtgggctt gtactcggtc atggtaagct agcttgggct gcaggtcgaa
     1381 aggcccggag atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag
     1441 aatgccgggc ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc
     1501 ccgcccccgg acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc
     1561 tattggccgc tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc
     1621 acgagactag tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg
     1681 gcggggggga acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag
     1741 aacggagccg gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt
     1801 gtgtagcgcc aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa
     1861 agcgcctccc ctacccggta gaattcgaac gctgacgtca tcaacccgct ccaaggaatc
     1921 gcgggcccag tgtcactagg cgggaacacc cagcgcgcgt gcgccctggc aggaagatgg
     1981 ctgtgaggga caggggagtg gcgccctgca atatttgcat gtcgctatgt gttctgggaa
     2041 atcaccataa acgtgaaatg tctttggatt tgggaatctt ataagttctg tatgagacca
     2101 cagatccccc attcaacatt cccttatatt caagagatat aagggaatgt tgaatggggg
     2161 atctaagctt atcgataccg tcgacctcga gggggggccc ggtaccagct tttgttccct
     2221 ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2281 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2341 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2401 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2461 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3061 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4021 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4381 gcgcacattt ccccgaaaag tgccac
//
