LOCUS       sequence-291408-        6637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsGIGYF2_1-718.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6637)
  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 6637)
  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-MS2-HA-C1-HsGIGYF2_1-718_AI
FEATURES             Location/Qualifiers
     source          1..6637
                     /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             1097..1123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3212..3223
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3425..3546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3462..3481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3516..3535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3553..4008)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3690..3711)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3902..3921
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4035..4139
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4141..4498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4141..4332
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4164..4184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4349..4484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4533..5327
                     /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             4533..5327
                     /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(4587..4606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5197..5216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5515..5534)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5559..5606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5935..6523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6424..6443
                     /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 caccatggct tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt
      721 gactgtcgcc ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct ctaactcgcg
      781 atcacaggct tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata
      841 caccatcaaa gtcgaggtgc ctaaagtggc aacccagact gttggtggtg aagagcttcc
      901 tgtagccgga tggagatctt acttaaatat ggaactaacc attccaattt tcgccacgaa
      961 ttccgactgc gagcttattg ttaaggcaat gcaaggtctc ctaaaagatg gaaacccgat
     1021 tccctcggcc atcgcggcca actccggcat ctacggaggt ggaggtggat ctggtggatc
     1081 tcctccacat atggcttacc catacgatgt tccagattac gctcctccat ctcgagctat
     1141 ggcagcggaa acgcagacac tgaactttgg gcctgaatgg ctccgagctc tgtccagtgg
     1201 tgggagtatt acatcccctc ctctttctcc agcattgccg aagtataaat tagcagatta
     1261 tcgttacggc agagaagaaa tgttagcact tttccttaaa gacaacaaga taccttcaga
     1321 ccttctggat aaagaatttc tgcctatcct ccaggaggaa ccccttccac cattggctct
     1381 ggtacccttt acagaagaag aacagagaaa cttttccatg tctgtaaata gtgctgctgt
     1441 cctgcgattg acaggacgag gaggaggagg aacagtggtg ggggctccta gaggtcgaag
     1501 ttcttcaaga gggcgaggca gaggcagagg tgaatgtggt ttctaccaaa gaagttttga
     1561 tgaagtagag ggtgtttttg gtcgaggagg tggcagagaa atgcatagat cgcagagctg
     1621 ggaggaaagg ggtgacagac gttttgaaaa accaggacga aaagatgtag ggagaccaaa
     1681 ttttgaggaa ggtggaccaa catcagtagg gagaaagcat gaatttatac gctcagaaag
     1741 tgaaaattgg cgcatcttta gagaggaaca aaatggagaa gatgaagatg gaggttggcg
     1801 actagctgga tcaaggaggg atggagagag gtggcgacct cacagtcctg atggccctcg
     1861 ttctgcaggc tggcgggaac acatggaacg acgtcggagg tttgagtttg attttcgaga
     1921 tagagatgat gaacggggtt accgaagggt tcgctctggc agtgggagca tagatgatga
     1981 cagggatagc ttgcccgaat ggtgcttaga ggatgctgaa gaagaaatgg gtacatttga
     2041 ctcatctgga gcattccttt ctctaaaaaa agtacagaaa gagcctattc cagaagagca
     2101 ggagatggac ttccggcctg tggacgaagg ggaggagtgc tctgactctg agggtagcca
     2161 taatgaagag gccaaagaac ccgataagac aaataagaaa gaaggagaga aaacagatag
     2221 agtaggagtt gaagctagtg aggaaactcc ccagacctca tcatcatctg ctagaccagg
     2281 tactccttca gaccatcagt ctcaggaagc atcacagttt gagaggaaag atgaaccaaa
     2341 aactgagcaa acggaaaaag ctgaagagga gactcggatg gaaaatagtc taccagccaa
     2401 agtgcccagc agaggggatg aaatggttgc tgatgtccag cagcccctgt cgcagattcc
     2461 ttcagataca gcctctcctc ttctcatact tccacctcct gttcccaatc ctagtcctac
     2521 tctccggcca gttgaaacac cagttgtagg tgctcctggt atgggcagtg tttccacaga
     2581 acctgatgat gaagaaggtc tcaaacattt ggagcagcaa gctgagaaaa tggtggctta
     2641 tctccaagac agtgcactag atgatgaaag attggcatca aaactgcaag agcacagagc
     2701 taaaggagtg tcgattccat tgatgcatga agcaatgcag aagtggtatt acaaagatcc
     2761 tcagggagaa attcaaggtc ccttcaataa tcaggagatg gcagaatggt ttcaggcggg
     2821 ctattttact atgtctttat tggtgaagag agcgtgtgat gaaagcttcc aacctcttgg
     2881 cgatatcatg aaaatgtggg gaagggttcc cttttctcca ggtccagctc cccctcctca
     2941 tatgggagag ctggaccagg aacgactgac caggcagcaa gaactcacag ccttatacca
     3001 gatgcagcac ctgcagtacc agcagttttt aatacaacaa caatatgcac aggttttggc
     3061 ccaacagcag aaagcagcac tgtcttccca gcagcagcag cagttggcac ttcttcttca
     3121 acagtttcag accttgaaga tgagaatatc tgatcagaac atcattccct cagtaactag
     3181 gtctgtgtcc gtgccagata ctggctctat ctgggagctt cagccaacag cttcacagcc
     3241 tacagtttgg gaaggtggta gtgtatggga tcttcctctg gacaccacga catgaggatc
     3301 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3361 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3421 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3541 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3601 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3661 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3721 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3781 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3841 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3901 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3961 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4021 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4081 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4141 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4321 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4381 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4441 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4501 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4561 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4621 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4681 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4741 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4801 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4861 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4921 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4981 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5041 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5101 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5161 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5221 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5281 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5341 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5401 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5461 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5521 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5581 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5641 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5701 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5761 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6061 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6601 ccctgattct gtggataacc gtattaccgc catgcat
//
