LOCUS       sequence-291397-        6808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsGIGYF2_1-718-del280-310.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6808)
  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 6808)
  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-EGFP-C1-HsGIGYF2_1-718-del280-310_AI
FEATURES             Location/Qualifiers
     source          1..6808
                     /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             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3383..3394
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3596..3717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3633..3652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3687..3706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3724..4179)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3861..3882)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4073..4092
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4206..4310
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4312..4669
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4312..4503
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4335..4355)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4520..4655
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4704..5498
                     /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             4704..5498
                     /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(4758..4777)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5368..5387
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5686..5705)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5730..5777
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6106..6694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6595..6614
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctatggcagc ggaaacgcag acactgaact ttgggcctga
     1441 atggctccga gctctgtcca gtggtgggag tattacatcc cctcctcttt ctccagcatt
     1501 gccgaagtat aaattagcag attatcgtta cggcagagaa gaaatgttag cacttttcct
     1561 taaagacaac aagatacctt cagaccttct ggataaagaa tttctgccta tcctccagga
     1621 ggaacccctt ccaccattgg ctctggtacc ctttacagaa gaagaacaga gaaacttttc
     1681 catgtctgta aatagtgctg ctgtcctgcg attgacagga cgaggaggag gaggaacagt
     1741 ggtgggggct cctagaggtc gaagttcttc aagagggcga ggcagaggca gaggtgaatg
     1801 tggtttctac caaagaagtt ttgatgaagt agagggtgtt tttggtcgag gaggtggcag
     1861 agaaatgcat agatcgcaga gctgggagga aaggggtgac agacgttttg aaaaaccagg
     1921 acgaaaagat gtagggagac caaattttga ggaaggtgga ccaacatcag tagggagaaa
     1981 gcatgaattt atacgctcag aaagtgaaaa ttggcgcatc tttagagagg aacaaaatgg
     2041 agaagatgaa gatggaggtt ggcgactagc tggatcaagg agggatggag agaggtggcg
     2101 acctcacagt cctgatggcc ctcgttctgc aggctggcgg gaacacatgg aacgacgtcg
     2161 gaggtttgag tttgattttc gagatagaga tgatgaacgg ggttaccgaa gggttcgctc
     2221 tggcagtggg agcatagata aagtacagaa agagcctatt ccagaagagc aggagatgga
     2281 cttccggcct gtggacgaag gggaggagtg ctctgactct gagggtagcc ataatgaaga
     2341 ggccaaagaa cccgataaga caaataagaa agaaggagag aaaacagata gagtaggagt
     2401 tgaagctagt gaggaaactc cccagacctc atcatcatct gctagaccag gtactccttc
     2461 agaccatcag tctcaggaag catcacagtt tgagaggaaa gatgaaccaa aaactgagca
     2521 aacggaaaaa gctgaagagg agactcggat ggaaaatagt ctaccagcca aagtgcccag
     2581 cagaggggat gaaatggttg ctgatgtcca gcagcccctg tcgcagattc cttcagatac
     2641 agcctctcct cttctcatac ttccacctcc tgttcccaat cctagtccta ctctccggcc
     2701 agttgaaaca ccagttgtag gtgctcctgg tatgggcagt gtttccacag aacctgatga
     2761 tgaagaaggt ctcaaacatt tggagcagca agctgagaaa atggtggctt atctccaaga
     2821 cagtgcacta gatgatgaaa gattggcatc aaaactgcaa gagcacagag ctaaaggagt
     2881 gtcgattcca ttgatgcatg aagcaatgca gaagtggtat tacaaagatc ctcagggaga
     2941 aattcaaggt cccttcaata atcaggagat ggcagaatgg tttcaggcgg gctattttac
     3001 tatgtcttta ttggtgaaga gagcgtgtga tgaaagcttc caacctcttg gcgatatcat
     3061 gaaaatgtgg ggaagggttc ccttttctcc aggtccagct ccccctcctc atatgggaga
     3121 gctggaccag gaacgactga ccaggcagca agaactcaca gccttatacc agatgcagca
     3181 cctgcagtac cagcagtttt taatacaaca acaatatgca caggttttgg cccaacagca
     3241 gaaagcagca ctgtcttccc agcagcagca gcagttggca cttcttcttc aacagtttca
     3301 gaccttgaag atgagaatat ctgatcagaa catcattccc tcagtaacta ggtctgtgtc
     3361 cgtgccagat actggctcta tctgggagct tcagccaaca gcttcacagc ctacagtttg
     3421 ggaaggtggt agtgtatggg atcttcctct ggacaccacg acatgaggat ccaccggatc
     3481 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3541 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3601 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3661 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     3721 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3781 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3841 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3901 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3961 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4021 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4081 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4141 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4321 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4441 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4501 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4561 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4621 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4681 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4741 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4801 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4861 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4921 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4981 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5041 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5101 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5161 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5221 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5281 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5341 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5401 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5461 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5521 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5581 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5641 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5701 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5761 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5821 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5881 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5941 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6001 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6241 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6721 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6781 tgtggataac cgtattaccg ccatgcat
//
