LOCUS       sequence-291391-        6791 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 6791)
  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 6791)
  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-HsGIGYF1_1-671_AI
FEATURES             Location/Qualifiers
     source          1..6791
                     /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"
     polyA_signal    3579..3700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3616..3635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3670..3689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3707..4162)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3844..3865)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4056..4075
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4189..4293
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4295..4652
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4295..4486
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4318..4338)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4503..4638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4687..5481
                     /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             4687..5481
                     /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(4741..4760)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5351..5370
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5669..5688)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5713..5760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6089..6677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6578..6597
                     /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 agagacactc aactttgggc ctgagtggct
     1441 cagggccctg tccgggggcg gcagcgtggc ctccccaccc ccgtcccctg ccatgcccaa
     1501 atacaagctg gctgactacc gttatgggcg agaggaaatg ctggctctct acgtcaagga
     1561 gaacaaggtc ccggaagagc tgcaggacaa ggagttcgcc gcggtgctgc aggacgagcc
     1621 actgcagccc ctggctctgg agccgctgac tgaggaggaa cagagaaact tctccctgtc
     1681 agtgaacagc gtggctgtgc tgaggctgat ggggaaaggg gctggccccc ccctggctgg
     1741 cacctcccga ggcaggggca gcacgcggag ccgaggccgc ggccgtggtg acagctgctt
     1801 ttaccaaaga agcatcgaag aaggcgatgg ggcctttgga cgaagccccc gggaaatcca
     1861 gcgcagccag agctgggatg acagaggcga gaggcggttt gagaagtcag caaggcggga
     1921 tggagcacga tgtggctttg aggagggagg ggctggccca aggaaggagc acgcccgctc
     1981 agacagcgag aactggcgct ccctacggga ggaacaggag gaggaggagg agggcagctg
     2041 gaggctcgga gcagggcccc ggcgagacgg cgaccgctgg cgctccgcca gccctgatgg
     2101 tggtccccgc tctgctggct ggcgggaaca tggggaacgg cggcgcaagt ttgaatttga
     2161 tttgcgaggg gatcgaggag ggtgtggtga agaggagggg cggggagggg gaggcagctc
     2221 tcacctgcgg cggtgccgag cgcctgaagg ctttgaggag gacaaggatg ggctcccaga
     2281 gtggtgcctg gacgatgagg atgaagaaat gggcaccttt gatgcctctg gggccttctt
     2341 gcctctcaag aagggcccca aggagcccat tcctgaggag caggagctgg acttccaagg
     2401 gttggaggag gaggaggaac cttccgaagg gctagaggag gaagggcctg aggcaggtgg
     2461 gaaagagctg accccactgc ctcctcagga ggagaagtcc agctccccat ccccactgcc
     2521 caccctgggc ccactctggg ggacaaacgg ggatggggac gaaactgcag agaaagagcc
     2581 cccagcggcc gaagatgata ttcgggggat ccagctgagt cccggggtgg gctcctctgc
     2641 tggcccaccc ggagatctgg aggatgatga aggcttgaag cacctgcagc aggaggcgga
     2701 gaagctggtg gcctccctgc aggacagctc cttggaggag gagcagttca cggctgccat
     2761 gcagacccag ggcctgcgcc actctgcagc cgccactgcc ctcccgctca gccatggggc
     2821 tgcccggaag tggttctaca aggacccaca gggcgagatc caaggcccct tcacgacaca
     2881 ggagatggca gagtggttcc aggccggcta cttttccatg tcactgctgg tgaagcgggg
     2941 ctgcgatgag ggcttccagc cgctgggcga ggtgatcaag atgtggggcc gcgtgccctt
     3001 tgccccaggg ccctcacctc ccccactgct gggaaacatg gaccaggagc ggctgaagaa
     3061 gcaacaggag ctggccgcgg cggccttgta ccagcagctg cagcaccagc agtttctcca
     3121 gctggtcagc agccgccagc tcccacagtg cgcgctccga gaaaaggcag ctctggggga
     3181 cctgacaccg ccaccaccgc cgccgccaca gcagcagcag cagcagctca cggcattcct
     3241 gcagcagctc caggcgctca aaccccccag aggcggggac cagaacctgc tcccgacgat
     3301 gagccggtcc ttgtcggtgc cagattcggg ccgcctctgg gacgtacata cctcagcctc
     3361 atcacagtca ggtggtgagg ccagtctttg ggacatacca attaactctt cgacttgaga
     3421 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     3481 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3541 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3601 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3661 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3721 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3781 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3841 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3901 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3961 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4021 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4081 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4141 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4201 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4261 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4321 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4381 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4441 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4501 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4561 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4621 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4681 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4741 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4801 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4861 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4921 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4981 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5041 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5101 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5161 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5221 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5281 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5341 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5401 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5461 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5521 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5581 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5641 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5701 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5761 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5821 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5881 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6781 ccgccatgca t
//
