LOCUS       sequence-291415-        5627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRck-L338AK342A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5627)
  AUTHORS   Peter D, Ruscica V, Bawankar P, Weber R, Helms S, Valkov E, Igreja 
            C, Izaurralde E
  TITLE     Molecular basis for GIGYF-Me31B complex assembly in 4EHP-mediated 
            translational repression.
  JOURNAL   Genes Dev. 2019 Oct 1;33(19-20):1355-1360. doi: 
            10.1101/gad.329219.119. Epub 2019 Aug 22.
  PUBMED    31439631
REFERENCE   2  (bases 1 to 5627)
  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: "Genes 
            Dev."; date: "2019-10-1"; volume: "33(19-20):1355-1360. doi"; pages:
            " 10.1101/gad.329219.119. Epub 2019 Aug 22"
COMMENT     Sequence Label: pT7-V5-SBP-C1-HsRck-L338AK342A_AI
FEATURES             Location/Qualifiers
     source          1..5627
                     /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             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    2415..2536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2452..2471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2506..2525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2543..2998)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2680..2701)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2892..2911
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3025..3129
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3131..3488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3131..3322
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3154..3174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3339..3474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3523..4317
                     /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             3523..4317
                     /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(3577..3596)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4187..4206
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4505..4524)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4549..4596
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4925..5513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5414..5433
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct atgggtctgt ccagtcaaaa tggtcagctg agaggccctg tgaaacccac
      901 tggtggccct ggaggagggg gcacacagac acagcaacag atgaaccagc tgaaaaacac
      961 caacacaatc aataatggca ctcagcagca agcacagagt atgaccacca ctattaaacc
     1021 tggtgatgac tggaaaaaga ctttaaaact ccctccaaag gatctaagaa tcaaaacttc
     1081 ggatgtgacc tccacaaaag gaaatgagtt tgaagattac tgtttgaaac gggagttact
     1141 gatgggaatt tttgaaatgg gctgggaaaa gccatctcct attcaggagg agagcattcc
     1201 cattgcttta tctggtaggg atatcttagc tagagcaaaa aatggaacag gcaagagcgg
     1261 tgcctacctc attcccttac ttgaacggct agacctgaag aaggacaata tacaagcaat
     1321 ggtgattgtt cccactagag aacttgctct acaggtcagt caaatttgca tccaggtcag
     1381 caaacacatg ggaggggcca aagtgatggc aaccacagga ggaaccaatt tacgagatga
     1441 cataatgagg cttgatgata cagtgcacgt ggtgattgct acccctggga gaatcctgga
     1501 tcttattaag aaaggagtag caaaggttga tcatgtccag atgatagtat tggatgaggc
     1561 agataagttg ctgtcacagg attttgtgca gataatggag gatattattc tcacgctacc
     1621 taaaaacagg cagattttac tatattccgc tactttccct cttagtgtac agaagttcat
     1681 gaattcccat ttgcagaaac cctatgagat taacctgatg gaggaactaa ctctgaaggg
     1741 agtaacccag tactacgcat atgtaactga gcgccaaaaa gtacactgcc tcaacacact
     1801 tttctccagg cttcagataa accagtcgat cattttctgt aactcctctc agcgagttga
     1861 agccctagcc aaggccattt ctcaactggg ttattcttgc ttctatattc atgctaaaat
     1921 gaggcaggaa catcgaaatc gtgtatttca tgatttccga aatggcttat gccgcaatct
     1981 tgtttgcact gatctgttta cccgaggtat tgatatacaa gctgtgaatg tggtaataaa
     2041 ctttgatttc ccaaagctgg cagagaccta tctccatcgt attggaagat caggtcgctt
     2101 tggtcatctt ggcttagcca tcaacttgat cacatatgat gatcgcttca acctgaaaag
     2161 tattgaggag cagctgggaa cagaaattaa acctattccg agcaacattg ataagagcct
     2221 gtatgtggca gaataccaca gcgagcctgt agaagatgag aaaccttaac aattctgcag
     2281 tcgacggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2341 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2401 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2521 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2581 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2641 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2701 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2761 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2821 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2881 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2941 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3001 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3061 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3121 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3421 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3481 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3541 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3601 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3661 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3721 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3781 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3841 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3901 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3961 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4021 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4081 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4141 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4201 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4261 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4321 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4381 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4441 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4501 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4561 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4621 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4681 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4741 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4861 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5041 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5101 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5161 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5221 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5281 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5341 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5401 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5461 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5521 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5581 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
