LOCUS       sequence-291243-        8812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsHelz_1051-1942-GSSG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8812)
  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 8812)
  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: pLIB-2STREP-HsHelz_1051-1942-GSSG-MBP_AG
FEATURES             Location/Qualifiers
     source          1..8812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4048..4081)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4129..4220
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4145..4162
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4313..4336
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4346..4369
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             7073..8167
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="TEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEK
                     FPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIA
                     YPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAAD
                     GGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETA
                     MTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLEN
                     YLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWY
                     AVRTAVINAASGRQTVDEALKDAQT"
     polyA_signal    8321..8455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8358..8377)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8412..8431
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8565..8730
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cagctttgtt tgtactatca acaggttgaa ctgctgatca
     4021 acagatcctc tacgcggccg cggtaccata acttcgtata gcatacatta tacgaagtta
     4081 tctggtttta ctagtatcga ttcgcgacct actccggaat attaatagat catggagata
     4141 attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg
     4201 taataaaaaa acctataaat attccggatt attcataccg tcccaccatc gggcgcggat
     4261 cccggtccga agcgcgcgga attcaaaggc ctacgtcgac atgggcagcg gctggagcca
     4321 cccccagttc gagaagggca gcggctggag ccacccccag ttcgagaagc ccattgctct
     4381 gtgctctatt ggaagatgca ggaaattttg ggaacggttt attgccctgt gtcatgaaaa
     4441 cagtagccta catggaatca cttttgaaca gatcaaagcc cagttagagg ctttagaact
     4501 aaagaagaca tatgtgttga atccgctggc acctgaattt atcccccggg ctctaagact
     4561 gcagcattca ggaagtacca acaaacagca gcaatcacca cccaagggga aaagtcttca
     4621 tcatacccag aatgatcact tccagaatga tggaattgtt cagcccaatc cttctgtact
     4681 tattggcaat cctattagag catatactcc tccaccccct cttggacctc acccaaattt
     4741 gggaaaatct ccaagccctg ttcaaagaat agatcctcac actgggacaa gtattcttta
     4801 tgtacctgct gtctatggag ggaatgtagt tatgtcggtg cctttacctg taccatggac
     4861 aggataccag ggtaggtttg cagttgatcc tcgaattatt acacatcagg cagcaatggc
     4921 ctataacatg aacctattac agacacatgg acgaggatct cctattcctt atggccttgg
     4981 acatcaccca cctgtcacca taggccagcc acaaaatcag catcaggaga aggatcaaca
     5041 tgagcaaaat cgaaatggta aaagtgatac aaataattcc ggacctgaaa ttaataagat
     5101 tcgaacacca gagaaaaagc caacagaacc aaaacaggtt gatttggaat caaatccaca
     5161 gaacagaagt cctgaatcac gtcctagtgt tgtttatccc agtaccaaat ttcctcgcaa
     5221 agataatctc aacccaagac acataaatct tccccttcct gctccccacg cacagtatgc
     5281 aatccctaat cgccactttc atccccttcc ccagctacca agaccaccct ttccaattcc
     5341 acagcagcac accttgttaa atcagcagca gaataatttg cctgaacaac caaatcagat
     5401 accacctcag ccaaatcagg tagtccagca gcaaagtcag ttgaatcagc agcctcagca
     5461 gccacctcct cagctttctc ctgcatatca ggcgggaccc aacaatgctt tttttaatag
     5521 tgcagttgct catcggccac agtctcctcc tgcagaagct gtaattccgg agcagcagcc
     5581 ccctcccatg ctgcaagaag gccacagtcc tctgagagcc attgcacaac ccggccccat
     5641 tcttccttca catctgaata gcttcattga tgagaacccc tcgggattac ctatagggga
     5701 ggctttagat cgtatacatg ggagtgtcgc tctggaaaca ttaaggcagc agcaggcacg
     5761 gttccagcag tggagcgagc atcatgcctt tctcagtcag ggcagcgttc catacccaca
     5821 ccatcaccat cctcacctcc agcatcttcc tcagccgccc ctgggattac atcagccgcc
     5881 agtgagggca gactggaagc tcaccagcag tgccgaagat gaagtggaga ccacatactc
     5941 aaggtttcaa gacttaatca gagaactgtc tcatcgtgat caaagtgaaa cacgggaact
     6001 agctgaaatg ccaccacctc aatcaagact tttgcaatat agacaagtac agagtagaag
     6061 cccaccagca gtcccatctc ccccatccag tacagaccac agtagccact tttctaactt
     6121 taatgataac agcagagaca ttgaagtagc cagcaaccca gcatttccac agcgcctccc
     6181 accccagata ttcaactcac ctttctcgtt gccatctgaa caccttgccc ctcctccctt
     6241 gaaatacctg gcacctgatg gagcatggac ttttgctaac ttgcaacaga atcacctaat
     6301 ggggccaggt tttccctatg gcctacctcc attgcctcac aggccaccgc agaacccttt
     6361 tgtacaaata cagaatcatc aacatgctat tggtcaagag ccatttcacc cattgtcatc
     6421 tcgaacagta tcttcttctt cgctccctag cttagaagag tatgagccca gaggacctgg
     6481 tcggcccttg taccaaagaa gaatctcatc tagctcagtt caaccttgtt ctgaagaagt
     6541 aagcactcct caagacagtc tggctcagtg taaagagctt caggaccaca gtaaccaatc
     6601 ttctttcaac ttttcatccc cggagtcctg ggtaaacacc acctcatcta ctccttatca
     6661 gaacattccg tgcaatggat ccagcaggac agctcagccc agagagttga tagcgccacc
     6721 caagactgtc aaaccccctg aggatcaact gaagtcggag aacctcgagg tgtccagttc
     6781 cttcaactac agtgtgctgc agcatcttgg ccagtttcca ccccttatgc ctaacaagca
     6841 gatcgcggag tcggccaata gcagtagccc ccagagctct gcggggggca agcccgccat
     6901 gtcctatgcc agcgctctgc gggcccctcc aaagcccagg ccccctcctg agcaggccaa
     6961 gaagagtagc gaccctctgt ctctcttcca ggaactgagc ctagggagct catctggcag
     7021 caatggcttt tactcatatt ttaaagggag ctcaggcggg agctcaggcg aaactgaaga
     7081 aggtaaactg gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg
     7141 taagaaattc gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga
     7201 agagaaattc ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca
     7261 cgaccgcttt ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc
     7321 gttccaggac aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat
     7381 tgcttacccg atcgctgttg aagcgttatc gctgatttat aacaaagacc tgctgccgaa
     7441 cccgccaaaa acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa
     7501 gagcgccctg atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga
     7561 cgggggttat gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga
     7621 taacgctggc gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat
     7681 gaatgcagac accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat
     7741 gaccatcaac ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt
     7801 aacggtactg ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc
     7861 aggtattaac gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct
     7921 gctgactgat gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct
     7981 gaagtcttac gaggaagagt tggcgaaaga tccacgtatt gccgccacga tggaaaacgc
     8041 ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg
     8101 tactgcggtg atcaacgccg ccagcggtcg tcagactgtc gatgaagccc ttaaagacgc
     8161 gcagacttaa tctagagcct gcagtctcga caagcttgtc gagaagtact agaggatcat
     8221 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     8281 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     8341 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     8401 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     8461 actgcttgag cctagaagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     8521 ctgccaccgc tgagcaataa ctatcataac ccctaggaga tccgaaccag ataagtgaaa
     8581 tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca
     8641 gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct
     8701 cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt
     8761 aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact tt
//
