LOCUS       sequence-420981-        6944 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian Expression of Hs4EHP-W95A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  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 6944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-V5-SBP-C1-Hs4EHP-W95A_AE
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(42..245)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        246..549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(724..1312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(804..823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1641..1688
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1713..1732
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1920..2714)
                     /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             complement(1920..2714)
                     /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(2031..2050)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2641..2660
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2749..3106)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2763..2898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2915..3106
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3063..3083
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3108..3212)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3239..3694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3326..3345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3536..3557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3701..3822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3712..3731)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3766..3785
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6710..6823)
                     /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"
     CDS             complement(6845..6886)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      6886..6895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6922..6941)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6923..6941)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tcgataagcc agtaagcagt gggttctcta gttagccaga gagctctgct tatatagacc
       61 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
      121 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
      181 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
      241 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
      301 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
      361 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
      421 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
      481 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
      541 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
      601 taataactaa tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1141 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1441 taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac
     1501 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     1561 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     1621 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
     1681 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     1741 ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     1801 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     1861 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     1921 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     1981 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     2041 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     2101 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     2161 acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg
     2221 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     2281 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     2341 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     2401 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     2461 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     2521 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     2581 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     2641 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     2701 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc
     2761 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     2821 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     2881 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     2941 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     3001 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     3061 acaccctaac tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt
     3121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3241 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3301 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3361 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3421 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3481 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3541 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3601 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3661 gcgaatttta acaaaatatt aacgcttaca atttacgcgt taagatacat tgatgagttt
     3721 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3781 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3841 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3901 tacaaatgtg gtatggctga ttatgatcag ttatctagat ccggtggatc ccgggcccgc
     3961 ggtaccgtcg actgcagaat tcctacgtga tgatcctggc gatggcttgc agggagggga
     4021 agtcgtcgtc ccgggcagca tgcagcgcct ggtggctgct ggcatcgcca tgcgcgagca
     4081 cctgctggca ggtggggcac acacggcagt ccaggcccgc ctgctgggtg gtggcctgcc
     4141 acgcgctctt cttgttcttc ttggacttgg tgctcagagg cttctcgcgg ttgcagaagt
     4201 ccgtgtgtgc agacaggagc tcctgctgct tggccgtgtc gggcagcagg accagcagct
     4261 cattaaagac cttctggaaa ttctccccca gcaggtcccg gcaactcttg taatactggg
     4321 ctgcggagat caggccctgt ctgaactccc ctgagtggct cttgaactcg ctgaagcggg
     4381 cctcgtcgct ctgcaggaag tccctgatgg actggatgag ctgaaggttc ctctcccgga
     4441 agttctcggg gactaggtac gcccgtgggg caggcagcag cctgggtgct tttgtggtgg
     4501 tggtggtggc ggggctgggg acacaggccg ggtgggggct aggcagaagg ccagagaagc
     4561 cagggggcgg cttgctgatt gggggcacca ggcccggcgg gggtggggga ggcgtgccct
     4621 tcaggagcac cacagcgctg aagcctgggg gcggcggcat ccggggtggg caggggccgc
     4681 cgagcgctgg gaagtcctcc tgtggcgtgg ggcaggggaa ggaccccagg ggccttggga
     4741 gcccaggggg ttccttgggg gcactccgag caggggccgg gccctccgtg tgtccattaa
     4801 cgacgacggc aactggcccc tcggctctgc tggcaggagc ttccggggcc tgcaaagccc
     4861 ctgggggaca ggtagcgggc gggggctgtg gcaatgtggt gcctggcttc tccgagccca
     4921 ctttcttctt cttcccaact ttagagggtt ggatggaggc cagccccagt gtggaggaga
     4981 cggagcccgt ggggcgtgtg ctcagaagct cctgcagggc cgggccgccg tcctcctcct
     5041 cctcctgtgt gaagggcggg ccgcccttcc tgccgcccct gctccccttc ccagccttcc
     5101 tggtgggctg gccgctgccg gtagcctgcg ccgagagtgg gggctgtgct accttcttgc
     5161 tgctactgct gctgttccag gcagagacaa ggctggtggg ggcggtgcca ggcttgggca
     5221 ccgaggacac cagggcgggg aagtcctcct cctggaaggc actcctgcct ctggcaggga
     5281 tggcgtacgg cagcgccaac cccacagggc ccggggttgc tgcagtggag caggaggagg
     5341 aagtggaggc agagaggctg gggaagtctt cgtccttgag cttcgggctg ggtggtggga
     5401 gggcgcctgg cacccccaca caccctgggg cggctgggcc tgtcaccgag aaggcttctt
     5461 ggcttacagg accatttgtc gaggtttcct tggggcctgg gccttcgccc tgagtccggg
     5521 gtgagcgccg ggggccacgg ggatcctcag gtccccgcgc cgccgcttct tccttttttg
     5581 gcctaccgcc ttcctcctgg tcttcgcttc ttcgggcctc ctcctgctgc tgtgcggcca
     5641 cggaggcccg gacagcagct gctacttctc ggtcctcttc ttcccttttg tacctccagc
     5701 ttcctcggcg gctctgctgg gctccgcgag tgccagcccg ggccactcgg ccctggcggc
     5761 tgtacctgtc cacctcctcg tagtcttcgc caccaacgac cccctcgttc cggcgcgagt
     5821 gccgtggcgc gtagctgaac tgcaggtcga tgtggcggtt ctggcgtgcc tcggcgcggc
     5881 tgcgactgtg gcaggccgtc ctgtgggcct tgaggtcgat ctcggtgcgg aaggcgtggg
     5941 tgaactgctc cgtgctgcag cggccttcct cacacagaaa gtgcttctcc cggaagtgct
     6001 cacgcaggta ggcatagtcg ctgtagtagt cctgggcccc gtccgagtcg cagaagtggc
     6061 agaagtagtg gtcgcggcgc aggtgcttaa gcagctcatc attgtccagg tagcgctcgt
     6121 cacagaactt gcagagcggg tgcccacggt gcgacgtgtc atcggggtca ccctgcatgc
     6181 gatgccgggc caggtccttg cgcgagtacc acttgcgctc atatgtgaag atctggaggt
     6241 gctggaggca cagacggcag cagaagagct catgctgcct ccgcatgtgc tgctccaggt
     6301 ccccgaagag gctgaaaggt ggcagctcgg ggcaccgcgg gcactcgtgc tgcagcagct
     6361 gcctgtacaa tgcgtacacc tttccatctg caaagtagat atcatatttc ttctcatgct
     6421 gcagctggtg gatggggatg gtggcaaagg caggaagctt cttcccaaag accacctggc
     6481 gcagctcctc gcggcacacg gcgcagtagc gctgctcgca gagcacccgc atcttggtag
     6541 agcagcggta gcacaccggg tggtcgcagc ggcccagcgc cgtggcctcc aggtctccgc
     6601 agcacagcac ggcgctcccg cctccccgct caggagctgc cgccgccgcc gcctccaggg
     6661 ccgcgcgccg cccctcggcg ccccccgccg ccgccatagc tcgagatctg ggctcccgct
     6721 ggccctgagg gtggtgctcc agcctggccc gcagctgctc cagctcgccg gccaggccct
     6781 ccaccacgtg gccgcccccc cagccggtgg tcttctcgtc catcgtaccg ctaccgcttc
     6841 cgggcgtaga atcgagaccg aggagagggt tagggatagg cttacccatg gtggcgaccg
     6901 gtagcgctag agcttgggtc tccctatagt gagtcgtatt aatt
//
