LOCUS       sequence-291254-        7657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsHelz_1-1050.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  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 7657)
  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_1-1050_AG
FEATURES             Location/Qualifiers
     source          1..7657
                     /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    4445..4566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4482..4501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4536..4555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4573..5028)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4710..4731)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4922..4941
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5055..5159
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5161..5518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5161..5352
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5184..5204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5369..5504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5553..6347
                     /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             5553..6347
                     /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(5607..5626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6217..6236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6535..6554)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6579..6626
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6955..7543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7444..7463
                     /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 tgattaatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4381 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4441 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4561 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4621 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4681 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4741 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4801 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4861 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4921 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4981 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5041 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5161 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5281 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5341 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5401 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5461 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5521 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5581 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5641 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5701 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5761 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5821 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5881 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5941 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6001 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6061 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6121 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6181 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6241 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6301 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6361 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6421 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6481 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6541 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6601 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6661 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6721 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6781 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7081 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     7141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7621 ccctgattct gtggataacc gtattaccgc catgcat
//
