LOCUS       sequence-357792-        7447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsHelz_1051-1942.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7447)
  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 7447)
  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-EGFP-HsHelz_1051-1942_AG
FEATURES             Location/Qualifiers
     source          1..7447
                     /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             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4235..4356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4272..4291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4326..4345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4363..4818)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4500..4521)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4712..4731
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4845..4949
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4951..5308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4951..5142
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4974..4994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5159..5294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5343..6137
                     /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             5343..6137
                     /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(5397..5416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6007..6026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6325..6344)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6369..6416
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6745..7333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7234..7253
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcggatcc
     1441 cattgctctg tgctctattg gaagatgcag gaaattttgg gaacggttta ttgccctgtg
     1501 tcatgaaaac agtagcctac atggaatcac ttttgaacag atcaaagccc agttagaggc
     1561 tttagaacta aagaagacat atgtgttgaa tccgctggca cctgaattta tcccccgggc
     1621 tctaagactg cagcattcag gaagtaccaa caaacagcag caatcaccac ccaaggggaa
     1681 aagtcttcat catacccaga atgatcactt ccagaatgat ggaattgttc agcccaatcc
     1741 ttctgtactt attggcaatc ctattagagc atatactcct ccaccccctc ttggacctca
     1801 cccaaatttg ggaaaatctc caagccctgt tcaaagaata gatcctcaca ctgggacaag
     1861 tattctttat gtacctgctg tctatggagg gaatgtagtt atgtcggtgc ctttacctgt
     1921 accatggaca ggataccagg gtaggtttgc agttgatcct cgaattatta cacatcaggc
     1981 agcaatggcc tataacatga acctattaca gacacatgga cgaggatctc ctattcctta
     2041 tggccttgga catcacccac ctgtcaccat aggccagcca caaaatcagc atcaggagaa
     2101 ggatcaacat gagcaaaatc gaaatggtaa aagtgataca aataattccg gacctgaaat
     2161 taataagatt cgaacaccag agaaaaagcc aacagaacca aaacaggttg atttggaatc
     2221 aaatccacag aacagaagtc ctgaatcacg tcctagtgtt gtttatccca gtaccaaatt
     2281 tcctcgcaaa gataatctca acccaagaca cataaatctt ccccttcctg ctccccacgc
     2341 acagtatgca atccctaatc gccactttca tccccttccc cagctaccaa gaccaccctt
     2401 tccaattcca cagcagcaca ccttgttaaa tcagcagcag aataatttgc ctgaacaacc
     2461 aaatcagata ccacctcagc caaatcaggt agtccagcag caaagtcagt tgaatcagca
     2521 gcctcagcag ccacctcctc agctttctcc tgcatatcag gcgggaccca acaatgcttt
     2581 ttttaatagt gcagttgctc atcggccaca gtctcctcct gcagaagctg taattccgga
     2641 gcagcagccc cctcccatgc tgcaagaagg ccacagtcct ctgagagcca ttgcacaacc
     2701 cggccccatt cttccttcac atctgaatag cttcattgat gagaacccct cgggattacc
     2761 tataggggag gctttagatc gtatacatgg gagtgtcgct ctggaaacat taaggcagca
     2821 gcaggcacgg ttccagcagt ggagcgagca tcatgccttt ctcagtcagg gcagcgttcc
     2881 atacccacac catcaccatc ctcacctcca gcatcttcct cagccgcccc tgggattaca
     2941 tcagccgcca gtgagggcag actggaagct caccagcagt gccgaagatg aagtggagac
     3001 cacatactca aggtttcaag acttaatcag agaactgtct catcgtgatc aaagtgaaac
     3061 acgggaacta gctgaaatgc caccacctca atcaagactt ttgcaatata gacaagtaca
     3121 gagtagaagc ccaccagcag tcccatctcc cccatccagt acagaccaca gtagccactt
     3181 ttctaacttt aatgataaca gcagagacat tgaagtagcc agcaacccag catttccaca
     3241 gcgcctccca ccccagatat tcaactcacc tttctcgttg ccatctgaac accttgcccc
     3301 tcctcccttg aaatacctgg cacctgatgg agcatggact tttgctaact tgcaacagaa
     3361 tcacctaatg gggccaggtt ttccctatgg cctacctcca ttgcctcaca ggccaccgca
     3421 gaaccctttt gtacaaatac agaatcatca acatgctatt ggtcaagagc catttcaccc
     3481 attgtcatct cgaacagtat cttcttcttc gctccctagc ttagaagagt atgagcccag
     3541 aggacctggt cggcccttgt accaaagaag aatctcatct agctcagttc aaccttgttc
     3601 tgaagaagta agcactcctc aagacagtct ggctcagtgt aaagagcttc aggaccacag
     3661 taaccaatct tctttcaact tttcatcccc ggagtcctgg gtaaacacca cctcatctac
     3721 tccttatcag aacattccgt gcaatggatc cagcaggaca gctcagccca gagagttgat
     3781 agcgccaccc aagactgtca aaccccctga ggatcaactg aagtcggaga acctcgaggt
     3841 gtccagttcc ttcaactaca gtgtgctgca gcatcttggc cagtttccac cccttatgcc
     3901 taacaagcag atcgcggagt cggccaatag cagtagcccc cagagctctg cggggggcaa
     3961 gcccgccatg tcctatgcca gcgctctgcg ggcccctcca aagcccaggc cccctcctga
     4021 gcaggccaag aagagtagcg accctctgtc tctcttccag gaactgagcc tagggagctc
     4081 atctggcagc aatggctttt actcatattt taaataatct agataactga tcataatcag
     4141 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4201 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     4261 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4321 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     4381 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4441 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4501 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4561 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4621 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4681 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4741 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4801 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4921 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4981 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5041 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5101 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5161 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5221 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5281 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     5341 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5401 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5461 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5521 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5581 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5641 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5701 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5761 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5821 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5881 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5941 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6001 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6061 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6121 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6181 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6241 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6301 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     6361 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6421 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6481 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6541 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6601 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6661 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6721 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6781 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6841 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6901 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6961 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7021 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7081 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7141 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7201 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7261 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7321 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7381 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7441 catgcat
//
