LOCUS       sequence-291406-        6527 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 6527)
  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 6527)
  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-MS2-HA-C1-HsGIGYF1_1-671_AI
FEATURES             Location/Qualifiers
     source          1..6527
                     /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             1097..1123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3315..3436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3352..3371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3406..3425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3443..3898)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3580..3601)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3792..3811
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3925..4029
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4031..4388
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4031..4222
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4054..4074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4239..4374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4423..5217
                     /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             4423..5217
                     /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(4477..4496)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5087..5106
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5405..5424)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5449..5496
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5825..6413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6314..6333
                     /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 caccatggct tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt
      721 gactgtcgcc ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct ctaactcgcg
      781 atcacaggct tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata
      841 caccatcaaa gtcgaggtgc ctaaagtggc aacccagact gttggtggtg aagagcttcc
      901 tgtagccgga tggagatctt acttaaatat ggaactaacc attccaattt tcgccacgaa
      961 ttccgactgc gagcttattg ttaaggcaat gcaaggtctc ctaaaagatg gaaacccgat
     1021 tccctcggcc atcgcggcca actccggcat ctacggaggt ggaggtggat ctggtggatc
     1081 tcctccacat atggcttacc catacgatgt tccagattac gctcctccat ctcgagctat
     1141 ggcagcagag acactcaact ttgggcctga gtggctcagg gccctgtccg ggggcggcag
     1201 cgtggcctcc ccacccccgt cccctgccat gcccaaatac aagctggctg actaccgtta
     1261 tgggcgagag gaaatgctgg ctctctacgt caaggagaac aaggtcccgg aagagctgca
     1321 ggacaaggag ttcgccgcgg tgctgcagga cgagccactg cagcccctgg ctctggagcc
     1381 gctgactgag gaggaacaga gaaacttctc cctgtcagtg aacagcgtgg ctgtgctgag
     1441 gctgatgggg aaaggggctg gcccccccct ggctggcacc tcccgaggca ggggcagcac
     1501 gcggagccga ggccgcggcc gtggtgacag ctgcttttac caaagaagca tcgaagaagg
     1561 cgatggggcc tttggacgaa gcccccggga aatccagcgc agccagagct gggatgacag
     1621 aggcgagagg cggtttgaga agtcagcaag gcgggatgga gcacgatgtg gctttgagga
     1681 gggaggggct ggcccaagga aggagcacgc ccgctcagac agcgagaact ggcgctccct
     1741 acgggaggaa caggaggagg aggaggaggg cagctggagg ctcggagcag ggccccggcg
     1801 agacggcgac cgctggcgct ccgccagccc tgatggtggt ccccgctctg ctggctggcg
     1861 ggaacatggg gaacggcggc gcaagtttga atttgatttg cgaggggatc gaggagggtg
     1921 tggtgaagag gaggggcggg gagggggagg cagctctcac ctgcggcggt gccgagcgcc
     1981 tgaaggcttt gaggaggaca aggatgggct cccagagtgg tgcctggacg atgaggatga
     2041 agaaatgggc acctttgatg cctctggggc cttcttgcct ctcaagaagg gccccaagga
     2101 gcccattcct gaggagcagg agctggactt ccaagggttg gaggaggagg aggaaccttc
     2161 cgaagggcta gaggaggaag ggcctgaggc aggtgggaaa gagctgaccc cactgcctcc
     2221 tcaggaggag aagtccagct ccccatcccc actgcccacc ctgggcccac tctgggggac
     2281 aaacggggat ggggacgaaa ctgcagagaa agagccccca gcggccgaag atgatattcg
     2341 ggggatccag ctgagtcccg gggtgggctc ctctgctggc ccacccggag atctggagga
     2401 tgatgaaggc ttgaagcacc tgcagcagga ggcggagaag ctggtggcct ccctgcagga
     2461 cagctccttg gaggaggagc agttcacggc tgccatgcag acccagggcc tgcgccactc
     2521 tgcagccgcc actgccctcc cgctcagcca tggggctgcc cggaagtggt tctacaagga
     2581 cccacagggc gagatccaag gccccttcac gacacaggag atggcagagt ggttccaggc
     2641 cggctacttt tccatgtcac tgctggtgaa gcggggctgc gatgagggct tccagccgct
     2701 gggcgaggtg atcaagatgt ggggccgcgt gccctttgcc ccagggccct cacctccccc
     2761 actgctggga aacatggacc aggagcggct gaagaagcaa caggagctgg ccgcggcggc
     2821 cttgtaccag cagctgcagc accagcagtt tctccagctg gtcagcagcc gccagctccc
     2881 acagtgcgcg ctccgagaaa aggcagctct gggggacctg acaccgccac caccgccgcc
     2941 gccacagcag cagcagcagc agctcacggc attcctgcag cagctccagg cgctcaaacc
     3001 ccccagaggc ggggaccaga acctgctccc gacgatgagc cggtccttgt cggtgccaga
     3061 ttcgggccgc ctctgggacg tacatacctc agcctcatca cagtcaggtg gtgaggccag
     3121 tctttgggac ataccaatta actcttcgac ttgagaattc tgcagtcgac ggtaccgcgg
     3181 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     3241 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3301 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3361 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3421 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3481 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3541 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3601 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3661 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3721 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3781 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3841 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3901 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3961 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4021 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4081 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4141 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4261 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4321 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4381 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4441 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4501 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4561 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4621 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4681 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4741 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4801 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4861 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4921 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4981 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5041 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5101 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5161 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5221 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5281 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5341 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5401 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5461 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5521 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5581 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5641 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5941 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6481 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
