LOCUS       sequence-426377-        7718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Hs4E-T.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7718)
  AUTHORS   Peter D, Weber R, Sandmeir F, Wohlbold L, Helms S, Bawankar P, 
            Valkov E, Igreja C, Izaurralde E
  TITLE     GIGYF1/2 proteins use auxiliary sequences to selectively bind to 
            4EHP and repress target mRNA expression.
  JOURNAL   Genes Dev. 2017 Jun 1;31(11):1147-1161. doi: 10.1101/gad.299420.117.
            Epub 2017 Jul 11.
  PUBMED    28698298
REFERENCE   2  (bases 1 to 7718)
  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: 
            "10.1101/gad.299420.117"; journalName: "Genes Dev"; date: 
            "2017-06-1- 1"; volume: "31"; issue: "11"; pages: "1147-1161"
COMMENT     Sequence Label: pT7-EGFP-C1-Hs4E-T_AF
FEATURES             Location/Qualifiers
     source          1..7718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /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(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4446..4567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4483..4502)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4537..4556
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4574..5029)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4711..4732)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4923..4942
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5056..5160
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5162..5519
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5162..5353
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5185..5205)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5370..5505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5554..6348
                     /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             5554..6348
                     /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(5608..5627)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6218..6237
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6536..6555)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6580..6627
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6956..7544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7445..7464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag ctcaagcttc gatggatagg agaagtatgg gtgaaacaga
     1381 aagtggagat gctttccttg acctgaagaa gcctcctgcc tccaaatgcc cccatcgcta
     1441 tacaaaagaa gaactcttgg atataaaaga actcccccat tccaaacaga ggccttcatg
     1501 cctttctgaa aaatatgaca gtgatggtgt ctgggaccct gagaagtggc atgcctctct
     1561 ctacccagct tcagggcgga gctcaccagt ggaaagtctg aagaaagagt tggatacaga
     1621 ccggccttcc ctggtgcgca ggatagtaga tccacgagag cgtgtgaaag aagatgactt
     1681 agatgttgtt ctcagccctc agagacggag ctttggaggg ggctgccacg tgacagccgc
     1741 tgttagctcc cggcgctcag gaagtccatt agagaaagat agtgatgggc ttcgtctgct
     1801 tggtggacgt aggattggca gtgggaggat aatctctgcc cggacctttg agaaggatca
     1861 ccgtcttagc gataaggacc tgcgggactt gagagacaga gaccgagaga gggacttcaa
     1921 ggacaagcgt ttcaggagag agtttggaga tagtaagcgt gtctttggtg agcgtagaag
     1981 aaatgattct tacacagaag aagaaccaga gtggttctct gctggaccca caagtcagtc
     2041 tgaaaccatc gaactgactg gctttgatga taagatacta gaagaagatc acaaagggag
     2101 aaaaagaaca aggcgacgga cagcctctgt gaaggaaggt atagtagagt gcaatggagg
     2161 agtggccgaa gaggatgaag tggaggtcat ccttgcacag gagcctgcgg ctgatcagga
     2221 agtgccaagg gatgctgtct tgcctgagca gtccccagga gactttgact ttaatgagtt
     2281 ctttaacctt gataaggtgc catgcttggc ttcgatgata gaagatgttt tgggagaagg
     2341 gtcagtctct gccagtcggt tcagtaggtg gttctctaac ccgagcagat caggaagccg
     2401 atccagcagt cttgggtcaa caccacatga agagctagag agacttgcag gtctggagca
     2461 agccatcctc tctcctggac agaactcggg gaattacttt gctcctatac cattggaaga
     2521 ccatgctgaa aataaagtgg atattttaga aatgctacag aaagccaaag tggatttgaa
     2581 acctcttctt tccagccttt ctgcaaataa agaaaaactt aaagaaagct cacattcagg
     2641 ggttgtgctt tcagtggagg aggtagaagc aggtctgaag ggcttgaagg ttgaccagca
     2701 agtgaagaat tcaactccct tcatggcaga acacctagaa gagaccttga gtgccgtaac
     2761 caacaatcga caactgaaga aagacggaga catgactgcg ttcaacaagc tagtgagcac
     2821 aatgaaggca agtgggactt tgccttctca gcccaaagtc agccgaaacc ttgaaagcca
     2881 tttgatgtcc cctgctgaga ttccaggcca gcctgtccct aagaacatcc tgcaggaact
     2941 tctgggtcaa ccagttcaga gacctgcttc ttccaatctt ctgagtggcc ttatggggag
     3001 cttggagcct acaacatctt tactgggcca aagagcaccc tctcctccct tgtcacaggt
     3061 gtttcaaact cgagcagcct cagctgacta ccttcgccca agaataccat caccaattgg
     3121 tttcacacca ggaccacagc agctactcgg agatccattc caaggcatgc gcaaacccat
     3181 gagccccatc acagcccaga tgagccagct ggagttgcaa caggcagctt tagaagggct
     3241 ggccttgcca catgaccttg ctgtacaggc agcaaacttc taccagcctg gttttggcaa
     3301 accacaggtg gacagaacca gagatggatt cagaaacagg caacagcgag tgaccaagtc
     3361 accagcaccc gtgcatcgag ggaattcctc ttcccctgcc cctgctgcct ccatcacaag
     3421 catgctttct ccttccttta cccctacctc agtgattcgt aagatgtacg agagcaaaga
     3481 gaaaagcaag gaggagccag catctggaaa agcagctctt ggtgacagta aagaggatac
     3541 tcagaaggcc agtgaagaaa acctcctgtc atccagctct gtacccagtg ccgatcgaga
     3601 ctcttctccc actacaaatt ccaaactgtc agcattacag aggtcttcgt gttccacccc
     3661 actgtcccag gccaaccgtt acaccaaaga acaagattat cgacctaaag caactgggag
     3721 aaaaacaccc accttggcat ccccagttcc tacaacacct tttctccgcc ctgtccacca
     3781 agttcccctt gtcccccatg tccctatggt taggcctgct caccagcttc acccagggtt
     3841 ggtacagagg atgctggccc agggagtaca tccacagcat cttccaagtt tgctccaaac
     3901 tggtgtgctt cctcctggga tggacttgag tcatttacag ggaatatctg gccccatcct
     3961 gggtcagccc ttttaccctt tacctgctgc tagtcaccct ctcttaaacc ctcgtcctgg
     4021 aacacctctg catctggcaa tggtgcaaca gcagctacag cgctcagttc tgcatcctcc
     4081 aggctctggt tcccatgcag cagctgtcag cgttcagaca acccctcaga acgtgcccag
     4141 ccggtcaggc ctgccccaca tgcactccca gctggagcat cgccccagcc agaggagcag
     4201 ctcccctgtg ggccttgcca aatggtttgg ctcagatgtg ctacagcaac ccctgccctc
     4261 catgcccgcc aaagttatca gtgtagatga attggaatac cgacagtgag ggcccgggat
     4321 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     4381 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4441 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4561 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4621 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4681 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4741 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4801 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4861 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4921 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4981 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5041 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5101 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5161 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5281 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5341 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5401 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5461 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     5521 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5581 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5641 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5701 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     5761 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5821 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5881 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5941 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6001 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6061 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     6121 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6181 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6241 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6301 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6361 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6421 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6481 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     6541 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6601 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     6661 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     6721 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6781 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6841 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6901 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6961 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7021 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7081 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7141 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7201 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7261 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7321 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7381 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7441 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7501 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7561 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7621 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     7681 acggggtcat tagttcatag cccatatatg gagttccg
//
