LOCUS       sequence-291407-        6434 bp    DNA     circular SYN 15-DEC-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 6434)
  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 6434)
  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-del286-316_AI
FEATURES             Location/Qualifiers
     source          1..6434
                     /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    3222..3343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3259..3278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3313..3332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3350..3805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3487..3508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3699..3718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3832..3936
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3938..4295
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3938..4129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3961..3981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4146..4281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4330..5124
                     /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             4330..5124
                     /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(4384..4403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4994..5013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5312..5331)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5356..5403
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5732..6320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6221..6240
                     /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 gagaagggcc ccaaggagcc cattcctgag gagcaggagc tggacttcca
     2041 agggttggag gaggaggagg aaccttccga agggctagag gaggaagggc ctgaggcagg
     2101 tgggaaagag ctgaccccac tgcctcctca ggaggagaag tccagctccc catccccact
     2161 gcccaccctg ggcccactct gggggacaaa cggggatggg gacgaaactg cagagaaaga
     2221 gcccccagcg gccgaagatg atattcgggg gatccagctg agtcccgggg tgggctcctc
     2281 tgctggccca cccggagatc tggaggatga tgaaggcttg aagcacctgc agcaggaggc
     2341 ggagaagctg gtggcctccc tgcaggacag ctccttggag gaggagcagt tcacggctgc
     2401 catgcagacc cagggcctgc gccactctgc agccgccact gccctcccgc tcagccatgg
     2461 ggctgcccgg aagtggttct acaaggaccc acagggcgag atccaaggcc ccttcacgac
     2521 acaggagatg gcagagtggt tccaggccgg ctacttttcc atgtcactgc tggtgaagcg
     2581 gggctgcgat gagggcttcc agccgctggg cgaggtgatc aagatgtggg gccgcgtgcc
     2641 ctttgcccca gggccctcac ctcccccact gctgggaaac atggaccagg agcggctgaa
     2701 gaagcaacag gagctggccg cggcggcctt gtaccagcag ctgcagcacc agcagtttct
     2761 ccagctggtc agcagccgcc agctcccaca gtgcgcgctc cgagaaaagg cagctctggg
     2821 ggacctgaca ccgccaccac cgccgccgcc acagcagcag cagcagcagc tcacggcatt
     2881 cctgcagcag ctccaggcgc tcaaaccccc cagaggcggg gaccagaacc tgctcccgac
     2941 gatgagccgg tccttgtcgg tgccagattc gggccgcctc tgggacgtac atacctcagc
     3001 ctcatcacag tcaggtggtg aggccagtct ttgggacata ccaattaact cttcgacttg
     3061 agaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca
     3121 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3181 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3361 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3421 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3481 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3541 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3601 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3661 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3721 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3781 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3841 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3901 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4021 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4201 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4261 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4321 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5161 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5221 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5281 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5341 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5401 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5461 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5521 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6361 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6421 ttaccgccat gcat
//
