LOCUS       sequence-290922-        4572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsHelz-shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4572)
  AUTHORS   Hanet A, Rasch F, Weber R, Ruscica V, Fauser M, Raisch T, 
            Kuzuoglu-Ozturk D, Chang CT, Bhandari D, Igreja C, Wohlbold L
  TITLE     HELZ directly interacts with CCR4-NOT and causes decay of bound 
            mRNAs.
  JOURNAL   Life Sci Alliance. 2019 Sep 30;2(5). pii: 2/5/e201900405. doi: 
            10.26508/lsa.201900405. Print 2019 Oct.
  PUBMED    31570513
REFERENCE   2  (bases 1 to 4572)
  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: "Life Sci 
            Alliance."; date: "2019-09-30"; volume: "2(5). pii: 2/5/e201900405. 
            doi"; pages: " 10.26508/lsa.201900405. Print 2019 Oct"
COMMENT     Sequence Label: pSUPERpuro-BglII-HsHelz_AC
FEATURES             Location/Qualifiers
     source          1..4572
                     /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     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        712..926
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     847..866
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(996..1012)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(997..1013)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        1014..1343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1194..1329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1440..2039
                     /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(1440..1459)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1936..1956
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2269..2350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2306..2325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2384..2404)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2384..2402)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2423..2439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2423..2439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2436..2458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2447..2463
                     /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(2471..2477,2478..2495,2496..2501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2516..2537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2654..2671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2825..3413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2905..2924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3584..4375,4376..4444))
                     /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     4207..4226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4445..4549)
                     /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 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gactagtgga tcccccgggc tgcaggaatt cgaacgctga
      721 cgtcatcaac ccgctccaag gaatcgcggg cccagtgtca ctaggcggga acacccagcg
      781 cgcgtgcgcc ctggcaggaa gatggctgtg agggacaggg gagtggcgcc ctgcaatatt
      841 tgcatgtcgc tatgtgttct gggaaatcac cataaacgtg aaatgtcttt ggatttggga
      901 atcttataag ttctgtatga gaccacagat cccccgcagt tgatcctcga attattcaag
      961 agataattcg aggatcaact gctttttaag cttatcgata ccgtcgacct cgagtgtgtc
     1021 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     1081 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1141 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     1201 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1261 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     1321 ttggaggcct aggcttttgc aaaaagctag cttgcatgcc tgcaggtcgg ccgccacgac
     1381 cggtgccgcc accatcccct gacccacgcc cctgacccct cacaaggaga cgaccttcca
     1441 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     1501 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     1561 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     1621 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     1681 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     1741 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     1801 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     1861 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     1921 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     1981 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     2041 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat ggctccgacc
     2101 gaagccaccc ggggcggccc cgccgacccc gcacccgccc ccgaggccca ccgactctag
     2161 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2221 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2281 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2341 ttttcactgc ctcgaggggg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     2401 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3241 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4561 gaaaagtgcc ac
//
