LOCUS       sequence-358331-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsGIGYF1_1-671.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  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 6245)
  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-HsGIGYF1_1-671_AI
FEATURES             Location/Qualifiers
     source          1..6245
                     /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    3033..3154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3070..3089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3124..3143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3161..3616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3298..3319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3510..3529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3643..3747
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3749..4106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3749..3940
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3772..3792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3957..4092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4141..4935
                     /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             4141..4935
                     /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(4195..4214)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4805..4824
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5123..5142)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5167..5214
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5543..6131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6032..6051
                     /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 cgagctatgg cagcagagac actcaacttt gggcctgagt ggctcagggc
      901 cctgtccggg ggcggcagcg tggcctcccc acccccgtcc cctgccatgc ccaaatacaa
      961 gctggctgac taccgttatg ggcgagagga aatgctggct ctctacgtca aggagaacaa
     1021 ggtcccggaa gagctgcagg acaaggagtt cgccgcggtg ctgcaggacg agccactgca
     1081 gcccctggct ctggagccgc tgactgagga ggaacagaga aacttctccc tgtcagtgaa
     1141 cagcgtggct gtgctgaggc tgatggggaa aggggctggc ccccccctgg ctggcacctc
     1201 ccgaggcagg ggcagcacgc ggagccgagg ccgcggccgt ggtgacagct gcttttacca
     1261 aagaagcatc gaagaaggcg atggggcctt tggacgaagc ccccgggaaa tccagcgcag
     1321 ccagagctgg gatgacagag gcgagaggcg gtttgagaag tcagcaaggc gggatggagc
     1381 acgatgtggc tttgaggagg gaggggctgg cccaaggaag gagcacgccc gctcagacag
     1441 cgagaactgg cgctccctac gggaggaaca ggaggaggag gaggagggca gctggaggct
     1501 cggagcaggg ccccggcgag acggcgaccg ctggcgctcc gccagccctg atggtggtcc
     1561 ccgctctgct ggctggcggg aacatgggga acggcggcgc aagtttgaat ttgatttgcg
     1621 aggggatcga ggagggtgtg gtgaagagga ggggcgggga gggggaggca gctctcacct
     1681 gcggcggtgc cgagcgcctg aaggctttga ggaggacaag gatgggctcc cagagtggtg
     1741 cctggacgat gaggatgaag aaatgggcac ctttgatgcc tctggggcct tcttgcctct
     1801 caagaagggc cccaaggagc ccattcctga ggagcaggag ctggacttcc aagggttgga
     1861 ggaggaggag gaaccttccg aagggctaga ggaggaaggg cctgaggcag gtgggaaaga
     1921 gctgacccca ctgcctcctc aggaggagaa gtccagctcc ccatccccac tgcccaccct
     1981 gggcccactc tgggggacaa acggggatgg ggacgaaact gcagagaaag agcccccagc
     2041 ggccgaagat gatattcggg ggatccagct gagtcccggg gtgggctcct ctgctggccc
     2101 acccggagat ctggaggatg atgaaggctt gaagcacctg cagcaggagg cggagaagct
     2161 ggtggcctcc ctgcaggaca gctccttgga ggaggagcag ttcacggctg ccatgcagac
     2221 ccagggcctg cgccactctg cagccgccac tgccctcccg ctcagccatg gggctgcccg
     2281 gaagtggttc tacaaggacc cacagggcga gatccaaggc cccttcacga cacaggagat
     2341 ggcagagtgg ttccaggccg gctacttttc catgtcactg ctggtgaagc ggggctgcga
     2401 tgagggcttc cagccgctgg gcgaggtgat caagatgtgg ggccgcgtgc cctttgcccc
     2461 agggccctca cctcccccac tgctgggaaa catggaccag gagcggctga agaagcaaca
     2521 ggagctggcc gcggcggcct tgtaccagca gctgcagcac cagcagtttc tccagctggt
     2581 cagcagccgc cagctcccac agtgcgcgct ccgagaaaag gcagctctgg gggacctgac
     2641 accgccacca ccgccgccgc cacagcagca gcagcagcag ctcacggcat tcctgcagca
     2701 gctccaggcg ctcaaacccc ccagaggcgg ggaccagaac ctgctcccga cgatgagccg
     2761 gtccttgtcg gtgccagatt cgggccgcct ctgggacgta catacctcag cctcatcaca
     2821 gtcaggtggt gaggccagtc tttgggacat accaattaac tcttcgactt gagaattctg
     2881 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     2941 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3001 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3121 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3181 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3241 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3301 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3361 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3421 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3481 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3541 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3601 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3661 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3721 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3781 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3841 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3901 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3961 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4021 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4081 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4141 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4201 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4261 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4321 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4381 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4441 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4501 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4561 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4621 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4681 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4741 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4801 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4861 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4921 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4981 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5041 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5101 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5161 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5221 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5281 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5341 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5401 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5461 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5521 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5581 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5641 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5701 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5761 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5821 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5881 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5941 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6001 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6061 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6241 tgcat
//
