LOCUS       sequence-291139-        8644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsGIGYF2-YYMLtoAAAA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8644)
  AUTHORS   Peter D, Weber R, Sandmeir F, Wohlbold L, Helms S, Bawankar P, 
            Valkov E, Igreja C, Izaurralde E
  TITLE     GIGYF1/2 proteins use auxiliary sequences to selectively bind to 
            4EHP and repress target mRNA expression.
  JOURNAL   Genes Dev. 2017 Jun 1;31(11):1147-1161. doi: 10.1101/gad.299420.117.
            Epub 2017 Jul 11.
  PUBMED    28698298
REFERENCE   2  (bases 1 to 8644)
  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"; doi: 
            "10.1101/gad.299420.117"; journalName: "Genes Dev"; date: 
            "2017-06-1- 1"; volume: "31"; issue: "11"; pages: "1147-1161"
COMMENT     Sequence Label: pT7-EGFP-C1-HsGIGYF2-YYMLtoAAAA_AF
FEATURES             Location/Qualifiers
     source          1..8644
                     /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             3476..3487
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    5432..5553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5469..5488)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5523..5542
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5560..6015)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5697..5718)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5909..5928
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6042..6146
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6148..6505
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6148..6339
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6171..6191)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6356..6491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6540..7334
                     /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             6540..7334
                     /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(6594..6613)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7204..7223
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7522..7541)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7566..7613
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7942..8530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8431..8450
                     /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 cgatggcagc ggaaacgcag acactgaact ttgggcctga
     1441 atggctccga gctctgtcca gtggtgggag tattacatcc cctcctcttt ctccagcatt
     1501 gccgaagtat aaattagcag atgcacgtgc cggcagagaa gaagcggcag 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 agcatagatg atgacaggga tagcttgccc gaatggtgct tagaggatgc
     2281 tgaagaagaa atgggtacat ttgactcatc tggagcattc ctttctctaa aaaaagtaca
     2341 gaaagagcct attccagaag agcaggagat ggacttccgg cctgtggacg aaggggagga
     2401 gtgctctgac tctgagggta gccataatga agaggccaaa gaacccgata agacaaataa
     2461 gaaagaagga gagaaaacag atagagtagg agttgaagct agtgaggaaa ctccccagac
     2521 ctcatcatca tctgctagac caggtactcc ttcagaccat cagtctcagg aagcatcaca
     2581 gtttgagagg aaagatgaac caaaaactga gcaaacggaa aaagctgaag aggagactcg
     2641 gatggaaaat agtctaccag ccaaagtgcc cagcagaggg gatgaaatgg ttgctgatgt
     2701 ccagcagccc ctgtcgcaga ttccttcaga tacagcctct cctcttctca tacttccacc
     2761 tcctgttccc aatcctagtc ctactctccg gccagttgaa acaccagttg taggtgctcc
     2821 tggtatgggc agtgtttcca cagaacctga tgatgaagaa ggtctcaaac atttggagca
     2881 gcaagctgag aaaatggtgg cttatctcca agacagtgca ctagatgatg aaagattggc
     2941 atcaaaactg caagagcaca gagctaaagg agtgtcgatt ccattgatgc atgaagcaat
     3001 gcagaagtgg tattacaaag atcctcaggg agaaattcaa ggtcccttca ataatcagga
     3061 gatggcagaa tggtttcagg cgggctattt tactatgtct ttattggtga agagagcgtg
     3121 tgatgaaagc ttccaacctc ttggcgatat catgaaaatg tggggaaggg ttcccttttc
     3181 tccaggtcca gctccccctc ctcatatggg agagctggac caggaacgac tgaccaggca
     3241 gcaagaactc acagccttat accagatgca gcacctgcag taccagcagt ttttaataca
     3301 acaacaatat gcacaggttt tggcccaaca gcagaaagca gcactgtctt cccagcagca
     3361 gcagcagttg gcacttcttc ttcaacagtt tcagaccttg aagatgagaa tatctgatca
     3421 gaacatcatt ccctcagtaa ctaggtctgt gtccgtgcca gatactggct ctatctggga
     3481 gcttcagcca acagcttcac agcctacagt ttgggaaggt ggtagtgtat gggatcttcc
     3541 tctggacacc acgacaccag gccctgccct ggaacagctt cagcagctag agaaggccaa
     3601 agctgcaaag ctagagcaag agagaagaga ggcagaaatg agggcaaaac gggaagagga
     3661 agagcgaaag aggcaggaag aactccgaag acaacaggag gaaattcttc ggcgacagca
     3721 ggaagaagaa aggaaaaggc gagaggaaga agaacttgcc cgaaggaaac aggaagaggc
     3781 tctgcgtcgc cagcgggagc aagaaattgc attaaggcga cagcgagaag aggaagaaag
     3841 acagcagcaa gaagaagctc ttagaagact ggaagagagg agaagagaag aggaagaaag
     3901 gcggaagcag gaagaattgt tacgcaaaca ggaagaggag gctgcaaaat gggcccggga
     3961 agaagaagaa gcccagcgtc gattagagga gaaccggctg cggatggaag aggaggcagc
     4021 cagactccgg catgaggaag aagaacggaa gagaaaggag ctggaggtcc agcggcagaa
     4081 ggagttaatg cgccagaggc agcagcagca agaggctctc cggaggttgc agcagcagca
     4141 gcagcaacaa cagctggcgc agatgaagct tccttcttct tcaacgtggg gccagcagtc
     4201 caatacaaca gcatgtcagt cccaggccac gctgtcgttg gctgaaatcc aaaaactaga
     4261 ggaagaacga gaacggcagc ttcgagaaga gcaaaggcgc cagcagaggg agttgatgaa
     4321 agctcttcag cagcagcagc agcagcaaca gcagaaactc tcaggttggg ggaatgtcag
     4381 caaaccttca ggtaccacga aatctcttct ggagatccag caggaagagg ccaggcaaat
     4441 gcaaaagcag cagcagcagc agcagcaaca ccagcaacca aacagagctc gtaacaatac
     4501 gcattccaac ctgcacacca gcattgggaa ttctgtttgg ggctctataa atactggtcc
     4561 tcctaaccag tgggcatctg acctagtcag tagtatttgg agtaatgctg acactaaaaa
     4621 ctccaacatg ggattctggg atgatgcagt gaaagaggtg ggacctagga attcaacaaa
     4681 taaaaataaa aacaacgcca gtctcagtaa atctgtaggt gtgtctaacc ggcagaataa
     4741 gaaagtagaa gaagaagaaa agttgctgaa gctctttcag ggagtaaata aagcccaaga
     4801 tggatttacg cagtggtgtg aacagatgct tcatgccctt aatacggcaa ataacttgga
     4861 tgttcccaca tttgtttctt tcctgaaaga agtagaatct ccttatgagg tccatgatta
     4921 tatcagggcc tatttaggag atacttctga ggccaaggag tttgccaagc agttccttga
     4981 gcgccgtgcc aaacagaaag ccaaccagca gcgtcagcag cagcagctgc cgcagcagca
     5041 gcagcagcag ccgccacagc agccgccaca gcagccacaa cagcaggact ctgtgtgggg
     5101 gatgaaccac agtacactcc attcagtatt tcagaccaat caaagcaaca accaacaatc
     5161 caattttgag gctgtgcaga gtggcaagaa gaagaaaaag cagaagatgg tccgagcaga
     5221 tcccagttta ttaggatttt cagtcaatgc atcatcggag cgactcaaca tgggtgaaat
     5281 cgagacgttg gatgactact gaggatccac cggatctaga taactgatca taatcagcca
     5341 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     5401 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5461 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5521 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     5581 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5641 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5701 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5761 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5821 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5881 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5941 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     6001 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6061 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6121 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     6181 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6241 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6301 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6361 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6421 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6481 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     6541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6721 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     7081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7321 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     7381 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7441 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     7501 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     7561 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     7621 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     7681 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7741 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7801 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7861 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7921 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7981 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8041 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     8101 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8161 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8221 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8281 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8341 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8401 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8461 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8521 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8581 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     8641 gcat
//
