LOCUS       sequence-291328-        8929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsHelz_1-1474.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8929)
  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 8929)
  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: pT7-MS2-HA-HsHelz_1-1474_AH
FEATURES             Location/Qualifiers
     source          1..8929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1097..1123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5717..5838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5754..5773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5808..5827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5845..6300)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5982..6003)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6194..6213
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6327..6431
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6433..6790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6433..6624
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6456..6476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6641..6776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6825..7619
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6825..7619
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6879..6898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7489..7508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7807..7826)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7851..7898
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8227..8815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8716..8735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggct tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt
      721 gactgtcgcc ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct ctaactcgcg
      781 atcacaggct tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata
      841 caccatcaaa gtcgaggtgc ctaaagtggc aacccagact gttggtggtg aagagcttcc
      901 tgtagccgga tggagatctt acttaaatat ggaactaacc attccaattt tcgccacgaa
      961 ttccgactgc gagcttattg ttaaggcaat gcaaggtctc ctaaaagatg gaaacccgat
     1021 tccctcggcc atcgcggcca actccggcat ctacggaggt ggaggtggat ctggtggatc
     1081 tcctccacat atggcttacc catacgatgt tccagattac gctcctccat ctcgagctca
     1141 agcttcgaat tctgcagtcg acggtaccgc ggatatggaa gacagaagag ctgaaaagtc
     1201 atgtgaacaa gcatgtgaat cacttaagag gcaggactat gaaatggccc tcaagcactg
     1261 cacagaggcc cttctttctc ttggccagta ctccatggca gacttcacag ggccttgtcc
     1321 attggaaata gaacgcatca aaatcgagag tcttctctac agaattgcct catttttgca
     1381 actgaaaaat tatgtgcaag ctgatgaaga ttgtagacat gtgctgggag aaggactggc
     1441 caagggagaa gatgcctttc gggcagtgct ttgctgcatg cagctgaaag ggaagctcca
     1501 acctgtatcc accattcttg ccaagtcact cacaggagag tccctgaatg ggatggtaac
     1561 aaaggatttg acaagactaa aaacacttct ctcagaaaca gagacagcaa ctagtaacgc
     1621 cctctctgga tatcacgtgg aagacttaga tgaggggtct tgtaatggtt ggcatttccg
     1681 cccaccacct aggggaatca caagcagcga ggaatatact ttgtgtaaaa gatttttaga
     1741 acaaggaatc tgtaggtatg gtgcccagtg tacttcagca cattcccagg aagaactagc
     1801 agaatggcag aaaagatatg cttcacggct gataaaattg aaacagcaaa atgagaataa
     1861 acagctctca ggcagttaca tggaaacctt gatagaaaag tggatgaatt cattgtctcc
     1921 tgagaaagtg cttagtgaat gtatagaagg agtaaaggta gagcacaatc ctgacctgtc
     1981 agttactgtc agcaccaaaa aatcccacca gacatggacc tttgctctca cttgtaagcc
     2041 tgcaagaatg ctgtatcgtg tagcattgct ttatgatgct catcgtcctc attttagtat
     2101 cattgcaata tctgccggag atagtactac ccaggtatca caagaagtcc cagaaaactg
     2161 tcaagaatgg ataggaggaa agatggccca aaatggatta gatcattacg tgtataaagt
     2221 cgggatagca tttaacacag aaatatttgg aacttttcgc caaaccatag ttttcgactt
     2281 tggattggaa ccagtactca tgcaaagagt aatgattgat gcagcttcta cagaagatct
     2341 cgaatacctg atgcatgcaa aacagcagct agtaaccaca gctaaacgtt gggattcttc
     2401 ctctaagact attatagatt ttgaacctaa tgaaactact gatttggaga agagccttct
     2461 tatcagatac caaattcccc tctctgctga ccagctattt actcagtccg ttttagacaa
     2521 atcattgacc aagagcaact atcagtcacg gttacatgac cttctttata ttgaggagat
     2581 agcccagtat aaagaaatca gcaagttcaa ccttaaagtg caattgcaga ttctggcaag
     2641 cttcatgctc actggtgttt ctggaggtgc aaagtatgct cagaatggac aactttttgg
     2701 tcgctttaag cttactgaaa cactttctga agatactttg gctggacgac tggtgatgac
     2761 caaagtcaat gctgtttatt tattaccagt ccctaaacag aagttagtac agacccaggg
     2821 aaccaaagag aaggtttatg aagctactat tgaagaaaaa acaaaggaat atatattttt
     2881 aaggctatct agggaatgct gtgaagaact taatcttcgg cctgactgtg acacacaggt
     2941 tgaacttcag tttcaattaa atcgattacc cctctgtgaa atgcactatg cactagacag
     3001 gatcaaggac aatggggttt tgtttccaga catcagtatg actcccacca taccatggag
     3061 tcctaacaga caatgggatg aacagttgga tcctcgacta aatgcaaaac agaaagaggc
     3121 tgttctggcc attaccactc cacttgcaat ccagctgccg cctgtgctta tcatcggacc
     3181 ctatgggaca ggcaaaacgt tcactctagc tcaggctgtc aaacatattc tgcagcaaca
     3241 ggagactagg attctcattt gcacccattc taatagtgct gctgatctct acataaagga
     3301 ttatttacat ccatatgtag aagcaggcaa tccccaggca agacctctca gggtatattt
     3361 cagaaatcgc tgggtaaaga ctgtccaccc agttgtgcat cagtactgtt tgatctcaag
     3421 cgcacattcc acctttcaga tgccccagaa agaagatatt cttaaacatc gagtggtggt
     3481 tgttaccttg aatacttccc agtacctctg tcagttggac cttgaacctg ggttttttac
     3541 acacattcta ttagatgaag ctgcccaggc catggagtgt gaaaccatta tgcctctagc
     3601 attagcaact caaaacactc ggattgtctt ggctggtgat cacatgcagc tcagtccttt
     3661 tgtttacagc gagtttgcca gggagagaaa ccttcacgtt tcattacttg accgactcta
     3721 tgagcattac cctgctgagt tcccatgtag gattctcctg tgtgagaact accgctccca
     3781 tgaagctatc atcaattata cctctgagct tttctatgag ggcaaactga tggccagtgg
     3841 gaagcagcca gcacacaaag atttctaccc actaactttc tttacagcac gaggagaaga
     3901 tgtacaagaa aaaaatagca cagcttttta taataatgca gaggtgtttg aagtggtgga
     3961 acgtgtagaa gagttaagaa ggaagtggcc agtagcgtgg gggaagttag atgatggcag
     4021 tattggtgtg gtgactccat atgctgatca agtgtttaga atacgtgctg aacttcgaaa
     4081 aaagagatta tctgatgtta atgtagaaag ggtgctaaat gttcaaggaa agcaattcag
     4141 agttttgttt cttagcacag tacgtacaag acatacttgt aaacataaac agacaccaat
     4201 taaaaagaaa gagcaacttc tggaagattc cacagaggac ttagattatg gttttttatc
     4261 taactacaag cttctcaata ctgccatcac aagagcacaa tccctggttg ctgtggtggg
     4321 tgatcccatt gctctgtgct ctattggaag atgcaggaaa ttttgggaac ggtttattgc
     4381 cctgtgtcat gaaaacagta gcctacatgg aatcactttt gaacagatca aagcccagtt
     4441 agaggcttta gaactaaaga agacatatgt gttgaatccg ctggcacctg aatttatccc
     4501 ccgggctcta agactgcagc attcaggaag taccaacaaa cagcagcaat caccacccaa
     4561 ggggaaaagt cttcatcata cccagaatga tcacttccag aatgatggaa ttgttcagcc
     4621 caatccttct gtacttattg gcaatcctat tagagcatat actcctccac cccctcttgg
     4681 acctcaccca aatttgggaa aatctccaag ccctgttcaa agaatagatc ctcacactgg
     4741 gacaagtatt ctttatgtac ctgctgtcta tggagggaat gtagttatgt cggtgccttt
     4801 acctgtacca tggacaggat accagggtag gtttgcagtt gatcctcgaa ttattacaca
     4861 tcaggcagca atggcctata acatgaacct attacagaca catggacgag gatctcctat
     4921 tccttatggc cttggacatc acccacctgt caccataggc cagccacaaa atcagcatca
     4981 ggagaaggat caacatgagc aaaatcgaaa tggtaaaagt gatacaaata attccggacc
     5041 tgaaattaat aagattcgaa caccagagaa aaagccaaca gaaccaaaac aggttgattt
     5101 ggaatcaaat ccacagaaca gaagtcctga atcacgtcct agtgttgttt atcccagtac
     5161 caaatttcct cgcaaagata atctcaaccc aagacacata aatcttcccc ttcctgctcc
     5221 ccacgcacag tatgcaatcc ctaatcgcca ctttcatccc cttccccagc taccaagacc
     5281 accctttcca attccacagc agcacacctt gttaaatcag cagcagaata atttgcctga
     5341 acaaccaaat cagataccac ctcagccaaa tcaggtagtc cagcagcaaa gtcagttgaa
     5401 tcagcagcct cagcagccac ctcctcagct ttctcctgca tatcaggcgg gacccaacaa
     5461 tgcttttttt aatagtgcag ttgctcatcg gccacagtct cctcctgcag aagctgtaat
     5521 tccggagcag cagccccctc ccatgctgca agaaggccac agtcctctga gagccattgc
     5581 acaacccggc cccatttaat ctagataact gatcataatc agccatacca catttgtaga
     5641 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5701 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5761 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5821 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5881 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5941 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6001 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6061 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6121 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6181 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6241 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6301 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6361 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6421 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6541 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6601 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6721 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6781 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6841 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6901 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6961 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     7021 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     7081 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     7141 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     7201 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     7261 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7321 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     7381 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7441 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7501 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7561 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7621 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7801 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7861 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7921 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7981 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     8041 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     8101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8341 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     8401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
