LOCUS       sequence-401043-        4501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Hs4EBP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4501)
  AUTHORS   Gruner S, Peter D, Weber R, Wohlbold L, Chung MY, Weichenrieder O, 
            Valkov E, Igreja C, Izaurralde E
  TITLE     The Structures of eIF4E-eIF4G Complexes Reveal an Extended Interface
            to Regulate Translation Initiation.
  JOURNAL   Mol Cell. 2016 Nov 3;64(3):467-479. doi: 
            10.1016/j.molcel.2016.09.020. Epub 2016 Oct 20.
  PUBMED    27773676
REFERENCE   2  (bases 1 to 4501)
  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: 
            "10.1016/j.molcel.2016.09.020"; journalName: "Mol Cell"; date: 
            "2016-11-3- 3"; volume: "64"; issue: "3"; pages: "467-479"
COMMENT     Sequence Label: pT7-V5-SBP-C1-Hs4EBP3_Z
FEATURES             Location/Qualifiers
     source          1..4501
                     /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             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    1289..1410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1326..1345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1380..1399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1417..1872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1554..1575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1766..1785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1899..2003
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2005..2362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2005..2196
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2028..2048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2213..2348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2397..3191
                     /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             2397..3191
                     /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(2451..2470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3061..3080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3379..3398)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3423..3470
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3799..4387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4288..4307
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctatgt caacgtccac tagctgcccg attcccgggg gccgggacca
      901 gctgcccgac tgctacagca ccacgccggg gggcacgcta tacgccacta cccccggagg
      961 caccaggatc atctacgacc gaaagttcct gctggagtgc aagaactcac ccattgcccg
     1021 gacacccccc tgctgcctcc ctcagattcc cggggtcaca actcctccaa cagcccctct
     1081 ctccaagctg gaggagctga aggagcagga gacagaggaa gagatacccg atgacgcaca
     1141 atttgaaatg gacatctaag gatccaccgg atctagataa ctgatcataa tcagccatac
     1201 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1261 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1321 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1381 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     1441 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1501 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1561 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1621 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1681 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1741 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1801 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1861 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1981 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2041 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2101 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2161 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2221 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2281 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2341 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2401 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2461 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2521 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2581 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2641 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2701 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2761 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2821 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2881 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2941 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3001 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3061 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3121 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3181 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3241 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3301 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3361 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3421 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3481 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3541 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3601 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3661 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3721 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4441 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     4501 t
//
