LOCUS       sequence-290999-        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Hs4EHP-S99N.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4876)
  AUTHORS   Peter D, Weber R, Sandmeir F, Wohlbold L, Helms S, Bawankar P, 
            Valkov E, Igreja C, Izaurralde E
  TITLE     GIGYF1/2 proteins use auxiliary sequences to selectively bind to 
            4EHP and repress target mRNA expression.
  JOURNAL   Genes Dev. 2017 Jun 1;31(11):1147-1161. doi: 10.1101/gad.299420.117.
            Epub 2017 Jul 11.
  PUBMED    28698298
REFERENCE   2  (bases 1 to 4876)
  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"; doi: 
            "10.1101/gad.299420.117"; journalName: "Genes Dev"; date: 
            "2017-06-1- 1"; volume: "31"; issue: "11"; pages: "1147-1161"
COMMENT     Sequence Label: pLNHA-Hs4EHP-S99N_AE
FEATURES             Location/Qualifiers
     source          1..4876
                     /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             665..730
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1664..1785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1701..1720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1755..1774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1792..2247)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1929..1950)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2141..2160
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2274..2378
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2380..2737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2380..2571
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2403..2423)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2588..2723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2772..3566
                     /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             2772..3566
                     /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(2826..2845)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3436..3455
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3754..3773)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3798..3845
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4174..4762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4663..4682
                     /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 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctatga acaacaagtt cgacgctttg aaagatgatg acagtgggga
      841 ccatgatcag aatgaagaaa acagcacaca gaaagatggt gagaaggaaa aaacggaacg
      901 agacaagaat cagagcagta gcaagagaaa ggctgttgtc cctggaccgg cagagcatcc
      961 cctgcagtac aactacactt tttggtactc caggagaacc cccggccgtc ccacgagctc
     1021 acagagctat gaacagaata tcaaacagat tggcaccttt gcctctgtgg agcagttctg
     1081 gaggttttat aaccacatgg tacgtcctgg ggacctgaca ggccacagtg acttccatct
     1141 cttcaaagaa ggaattaaac ccatgtggga ggatgatgca aataaaaatg gtggcaagtg
     1201 gattattcgg ctgcggaagg gcttggcctc ccgttgctgg gagaatctca ttttggccat
     1261 gctgggggaa cagttcatgg ttggggagga gatctgtggg gctgtggtgt ctgtccgctt
     1321 tcaggaagac attatttcaa tatggaataa gactgccagt gaccaagcaa ccacagcccg
     1381 aatccgggac acacttcggc gagtgcttaa cctacctccc aacaccatta tggaatacaa
     1441 aactcacacc gacagcatca aaatgccagg caggctgggc ccccaaaggc tcctttttca
     1501 aaacctctgg aagccgcggt tgaatgtgcc atgaggatcc accggatcta gataactgat
     1561 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1621 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1681 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1741 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     1801 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1861 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     1921 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1981 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2041 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2101 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2161 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2221 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2281 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2341 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2401 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2461 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2521 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2581 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2641 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2701 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     2761 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2821 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2881 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2941 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3001 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3061 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3121 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3181 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3241 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3301 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3361 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3421 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3481 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3541 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3601 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3661 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3721 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     3781 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     3841 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3901 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3961 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4021 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4081 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4321 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4741 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4801 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4861 tattaccgcc atgcat
//
