LOCUS       sequence-489477-        7337 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Mammalian Expression of HsGIGYF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  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 7337)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 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_AI
FEATURES             Location/Qualifiers
     source          1..7337
                     /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    4125..4246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4162..4181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4216..4235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4253..4708)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4390..4411)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4602..4621
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4735..4839
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4841..5198
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4841..5032
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4864..4884)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5049..5184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5233..6027
                     /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             5233..6027
                     /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(5287..5306)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5897..5916
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6215..6234)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6259..6306
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6635..7223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7124..7143
                     /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 tcttcgactc agggtccaat
     2881 tctagaacaa ctccagctgc aacataaatt ccaggagcgc agagaagtgg agctcagggc
     2941 gaagcgggag gaagaggaac gcaagcgtcg agaggagaag cgccgccagc agcagcagga
     3001 ggagcagaag cggcggcagg aggaggaaga gctgtttcgg cgcaagcacg tgcggcagca
     3061 ggagctattg ctgaagttgc tacagcagca gcaggcggtc cctgtgcccc ccgcacccag
     3121 ctccccgccc ccactctggg ctggcctggc caagcagggg ctgtccatga agacgctcct
     3181 ggagttgcag ctggagggcg agcggcagct gcacaaacag cccccacctc gggagccagc
     3241 tcgggcccag gcccccaacc accgagtgca gcttgggggc ctgggcactg cccccctgaa
     3301 ccagtgggtg tctgaggctg ggccactgtg gggcgggcca gacaagagtg ggggcggcag
     3361 cagcggcctg gggctctggg aggacacccc caagagcggc gggagcctgg tccgtggcct
     3421 cggcctgaag aacagccgga gcagcccatc tctcagtgac tcatacagcc acctatcggg
     3481 tcggcccatt cgcaaaaaga cggaggaaga agagaagctg ctgaagctgc tgcagggcat
     3541 tcccaggccc caggacggct tcacccagtg gtgcgagcag atgctgcaca cgctgagcgc
     3601 cacgggcagc ctggacgtgc ccatggctgt agcgatcctc aaggaggtgg aatcccccta
     3661 tgatgtccac gattatatcc gttcctgcct gggggacacg ctggaagcca aagaatttgc
     3721 caaacaattc ctggagcgga gggccaagca gaaagccagc cagcagcggc agcagcagca
     3781 ggaggcatgg ctgagcagcg cctcgctgca gacggccttc caggccaacc acagcaccaa
     3841 actcggcccc ggggagggca gcaaggccaa gaggcgggca ctgatgctgc actcagaccc
     3901 cagcatcctg gggtactccc tgcacggatc ttctggtgag atcgagagcg tggatgacta
     3961 ctgagaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     4021 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4081 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4141 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4201 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     4261 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4321 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4381 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4441 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4501 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4561 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4621 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4681 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4801 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4861 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4921 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5041 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5101 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5161 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5221 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5281 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5341 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5401 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5461 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5521 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5581 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5641 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5701 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5761 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5821 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5881 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5941 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6001 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6061 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6121 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6181 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6241 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6301 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6361 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6421 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6481 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6541 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6601 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6661 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6721 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6781 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6841 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6901 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6961 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7021 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7081 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7141 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7201 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7261 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7321 gtattaccgc catgcat
//
