LOCUS       sequence-291392-        6698 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsGIGYF1_1-671-del286-316.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6698)
  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 6698)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-del286-316_AI
FEATURES             Location/Qualifiers
     source          1..6698
                     /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    3486..3607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3523..3542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3577..3596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3614..4069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3772)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3963..3982
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4096..4200
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4202..4559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4202..4393
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4225..4245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4410..4545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4594..5388
                     /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             4594..5388
                     /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(4648..4667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5258..5277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5576..5595)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5620..5667
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5996..6584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6485..6504
                     /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 ctttgagaag ggccccaagg agcccattcc
     2281 tgaggagcag gagctggact tccaagggtt ggaggaggag gaggaacctt ccgaagggct
     2341 agaggaggaa gggcctgagg caggtgggaa agagctgacc ccactgcctc ctcaggagga
     2401 gaagtccagc tccccatccc cactgcccac cctgggccca ctctggggga caaacgggga
     2461 tggggacgaa actgcagaga aagagccccc agcggccgaa gatgatattc gggggatcca
     2521 gctgagtccc ggggtgggct cctctgctgg cccacccgga gatctggagg atgatgaagg
     2581 cttgaagcac ctgcagcagg aggcggagaa gctggtggcc tccctgcagg acagctcctt
     2641 ggaggaggag cagttcacgg ctgccatgca gacccagggc ctgcgccact ctgcagccgc
     2701 cactgccctc ccgctcagcc atggggctgc ccggaagtgg ttctacaagg acccacaggg
     2761 cgagatccaa ggccccttca cgacacagga gatggcagag tggttccagg ccggctactt
     2821 ttccatgtca ctgctggtga agcggggctg cgatgagggc ttccagccgc tgggcgaggt
     2881 gatcaagatg tggggccgcg tgccctttgc cccagggccc tcacctcccc cactgctggg
     2941 aaacatggac caggagcggc tgaagaagca acaggagctg gccgcggcgg ccttgtacca
     3001 gcagctgcag caccagcagt ttctccagct ggtcagcagc cgccagctcc cacagtgcgc
     3061 gctccgagaa aaggcagctc tgggggacct gacaccgcca ccaccgccgc cgccacagca
     3121 gcagcagcag cagctcacgg cattcctgca gcagctccag gcgctcaaac cccccagagg
     3181 cggggaccag aacctgctcc cgacgatgag ccggtccttg tcggtgccag attcgggccg
     3241 cctctgggac gtacatacct cagcctcatc acagtcaggt ggtgaggcca gtctttggga
     3301 cataccaatt aactcttcga cttgagaatt ctgcagtcga cggtaccgcg ggcccgggat
     3361 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     3421 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3481 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3541 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3601 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3661 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3721 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3781 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3841 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3901 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3961 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4021 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4081 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4141 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4201 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4561 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4621 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4681 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4741 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4801 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4861 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4921 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4981 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5041 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5101 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5161 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5221 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5281 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5341 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5401 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5461 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5521 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5581 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5641 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5701 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5761 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5821 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6121 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6661 cccctgattc tgtggataac cgtattaccg ccatgcat
//
