LOCUS       sequence-291251-        7921 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 7921)
  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 7921)
  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_1-1050_AG
FEATURES             Location/Qualifiers
     source          1..7921
                     /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    4709..4830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4746..4765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4800..4819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4837..5292)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4974..4995)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5186..5205
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5319..5423
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5425..5782
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5425..5616
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5448..5468)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5633..5768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5817..6611
                     /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             5817..6611
                     /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(5871..5890)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6481..6500
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6799..6818)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6843..6890
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7219..7807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7708..7727
                     /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 ccgcggatat
     1441 ggaagacaga agagctgaaa agtcatgtga acaagcatgt gaatcactta agaggcagga
     1501 ctatgaaatg gccctcaagc actgcacaga ggcccttctt tctcttggcc agtactccat
     1561 ggcagacttc acagggcctt gtccattgga aatagaacgc atcaaaatcg agagtcttct
     1621 ctacagaatt gcctcatttt tgcaactgaa aaattatgtg caagctgatg aagattgtag
     1681 acatgtgctg ggagaaggac tggccaaggg agaagatgcc tttcgggcag tgctttgctg
     1741 catgcagctg aaagggaagc tccaacctgt atccaccatt cttgccaagt cactcacagg
     1801 agagtccctg aatgggatgg taacaaagga tttgacaaga ctaaaaacac ttctctcaga
     1861 aacagagaca gcaactagta acgccctctc tggatatcac gtggaagact tagatgaggg
     1921 gtcttgtaat ggttggcatt tccgcccacc acctagggga atcacaagca gcgaggaata
     1981 tactttgtgt aaaagatttt tagaacaagg aatctgtagg tatggtgccc agtgtacttc
     2041 agcacattcc caggaagaac tagcagaatg gcagaaaaga tatgcttcac ggctgataaa
     2101 attgaaacag caaaatgaga ataaacagct ctcaggcagt tacatggaaa ccttgataga
     2161 aaagtggatg aattcattgt ctcctgagaa agtgcttagt gaatgtatag aaggagtaaa
     2221 ggtagagcac aatcctgacc tgtcagttac tgtcagcacc aaaaaatccc accagacatg
     2281 gacctttgct ctcacttgta agcctgcaag aatgctgtat cgtgtagcat tgctttatga
     2341 tgctcatcgt cctcatttta gtatcattgc aatatctgcc ggagatagta ctacccaggt
     2401 atcacaagaa gtcccagaaa actgtcaaga atggatagga ggaaagatgg cccaaaatgg
     2461 attagatcat tacgtgtata aagtcgggat agcatttaac acagaaatat ttggaacttt
     2521 tcgccaaacc atagttttcg actttggatt ggaaccagta ctcatgcaaa gagtaatgat
     2581 tgatgcagct tctacagaag atctcgaata cctgatgcat gcaaaacagc agctagtaac
     2641 cacagctaaa cgttgggatt cttcctctaa gactattata gattttgaac ctaatgaaac
     2701 tactgatttg gagaagagcc ttcttatcag ataccaaatt cccctctctg ctgaccagct
     2761 atttactcag tccgttttag acaaatcatt gaccaagagc aactatcagt cacggttaca
     2821 tgaccttctt tatattgagg agatagccca gtataaagaa atcagcaagt tcaaccttaa
     2881 agtgcaattg cagattctgg caagcttcat gctcactggt gtttctggag gtgcaaagta
     2941 tgctcagaat ggacaacttt ttggtcgctt taagcttact gaaacacttt ctgaagatac
     3001 tttggctgga cgactggtga tgaccaaagt caatgctgtt tatttattac cagtccctaa
     3061 acagaagtta gtacagaccc agggaaccaa agagaaggtt tatgaagcta ctattgaaga
     3121 aaaaacaaag gaatatatat ttttaaggct atctagggaa tgctgtgaag aacttaatct
     3181 tcggcctgac tgtgacacac aggttgaact tcagtttcaa ttaaatcgat tacccctctg
     3241 tgaaatgcac tatgcactag acaggatcaa ggacaatggg gttttgtttc cagacatcag
     3301 tatgactccc accataccat ggagtcctaa cagacaatgg gatgaacagt tggatcctcg
     3361 actaaatgca aaacagaaag aggctgttct ggccattacc actccacttg caatccagct
     3421 gccgcctgtg cttatcatcg gaccctatgg gacaggcaaa acgttcactc tagctcaggc
     3481 tgtcaaacat attctgcagc aacaggagac taggattctc atttgcaccc attctaatag
     3541 tgctgctgat ctctacataa aggattattt acatccatat gtagaagcag gcaatcccca
     3601 ggcaagacct ctcagggtat atttcagaaa tcgctgggta aagactgtcc acccagttgt
     3661 gcatcagtac tgtttgatct caagcgcaca ttccaccttt cagatgcccc agaaagaaga
     3721 tattcttaaa catcgagtgg tggttgttac cttgaatact tcccagtacc tctgtcagtt
     3781 ggaccttgaa cctgggtttt ttacacacat tctattagat gaagctgccc aggccatgga
     3841 gtgtgaaacc attatgcctc tagcattagc aactcaaaac actcggattg tcttggctgg
     3901 tgatcacatg cagctcagtc cttttgttta cagcgagttt gccagggaga gaaaccttca
     3961 cgtttcatta cttgaccgac tctatgagca ttaccctgct gagttcccat gtaggattct
     4021 cctgtgtgag aactaccgct cccatgaagc tatcatcaat tatacctctg agcttttcta
     4081 tgagggcaaa ctgatggcca gtgggaagca gccagcacac aaagatttct acccactaac
     4141 tttctttaca gcacgaggag aagatgtaca agaaaaaaat agcacagctt tttataataa
     4201 tgcagaggtg tttgaagtgg tggaacgtgt agaagagtta agaaggaagt ggccagtagc
     4261 gtgggggaag ttagatgatg gcagtattgg tgtggtgact ccatatgctg atcaagtgtt
     4321 tagaatacgt gctgaacttc gaaaaaagag attatctgat gttaatgtag aaagggtgct
     4381 aaatgttcaa ggaaagcaat tcagagtttt gtttcttagc acagtacgta caagacatac
     4441 ttgtaaacat aaacagacac caattaaaaa gaaagagcaa cttctggaag attccacaga
     4501 ggacttagat tatggttttt tatctaacta caagcttctc aatactgcca tcacaagagc
     4561 acaatccctg gttgctgtgg tgggtgatta atctagataa ctgatcataa tcagccatac
     4621 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4681 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4801 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     4861 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4921 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4981 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5041 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5101 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5161 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5221 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5281 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5401 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     5461 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5521 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5581 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5641 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5701 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5761 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5821 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5881 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5941 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     6001 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6061 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6121 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6181 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6241 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6301 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     6361 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6421 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6481 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6541 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6601 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6661 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6721 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6781 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6841 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6901 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6961 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     7021 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     7081 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7141 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7321 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     7381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     7921 t
//
