LOCUS       sequence-291013-        4967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of Hs4EHP_52-234opt-R103LE149L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4967)
  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 4967)
  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: pnYC-CvH-Hs4EHP_52-234opt-R103LE149L_AE
FEATURES             Location/Qualifiers
     source          1..4967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2293..2312
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2401..2420
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2401..2419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2420..2444
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2459..2481
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3053..3073
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3101..3118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3199..3217)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3213..3260
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3435..3440,3441..3457,3458..3463))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3575..4120
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4021..4040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4237..4254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4646..4748
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4815..4834)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
       61 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
      121 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
      181 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
      241 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
      301 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
      361 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
      421 agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtg
      481 ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac
      541 ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc
      601 ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt
      661 gctcaggctc tccccgtgga ggtaataatt gacgatatga tcatttattc tgcctcccag
      721 agcctgataa aaacggttag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      781 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgcttgt ttcggcgtgg
      841 gtatggtggc aggccccgtg gccgggggac tgttgggcgc tgccggcacc tgtcctacga
      901 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
      961 ggaaggagct accggacagc ggtgcggact gttgtaactc agaataagaa atgaggccgc
     1021 tcatggcgtt gactctcagt catagtatcg tggtatcacc ggttggttcc actctctgtt
     1081 gcgggcaact tcagcagcac gtaggggact tccgcgtttc cagactttac gaaacacgga
     1141 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     1201 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     1261 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     1321 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     1381 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     1441 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     1501 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     1561 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccagtgat
     1621 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     1681 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     1741 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     1801 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     1861 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     1921 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     1981 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     2041 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg
     2101 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     2161 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     2221 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     2281 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     2341 gatgccggcc acgatgcgtc cggcgtagag gatcagacta agatctcgat cccgcgaaat
     2401 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
     2461 tgtttaactt taagaaggag atatacatat gctcgaggca gaacatccgc tgcagtataa
     2521 ctataccttt tggtatagcc gtcgtacacc gggtcgtccg accagcagcc agagctatga
     2581 acagaacatt aaacaaattg gcacctttgc aagcgtggaa cagttttggc gtttttatag
     2641 ccatatggtt ctaccgggtg atctgacagg tcatagcgat tttcacctgt ttaaagaagg
     2701 cattaaaccg atgtgggaag atgatgcaaa caaaaatggt ggcaaatgga ttattcgtct
     2761 gcgtaaaggt ctggcaagcc gttgttggct aaatctgatt ctggcaatgc tgggtgaaca
     2821 gtttatggtt ggtgaagaaa tttgtggtgc agttgttagc gttcgttttc aagaagatat
     2881 tatcagcatc tggaataaaa ccgcaagcga tcaggcaacc accgcacgta ttcgtgatac
     2941 cctgcgtcgt gttctgaatc tgcctccgaa taccattatg gaatacaaaa cccataccga
     3001 tagcatcaaa atgcctggtc gtctgggtcc gcagcgtctg ctgtttggat ccgaaaacct
     3061 ttacttccag ggcagcggcg ccatgggcag cggcagcggc caccaccacc accaccacgg
     3121 tacctaatag tctagagcta gccctaggag atccggctgc taacaaagcc cgaaaggaag
     3181 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3241 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
     3301 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
     3361 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     3421 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattacaact
     3481 tatatcgtat ggggctgact tcaggtgcta catttgaaga gataaattgc actgaaatca
     3541 agaaatattt tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
     3601 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
     3661 ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc
     3721 tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca
     3781 gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta
     3841 ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag
     3901 cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag
     3961 cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg
     4021 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca
     4081 gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct
     4141 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
     4201 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     4261 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
     4321 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccc
     4381 ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcag tagctgaaca
     4441 ggagggacag ctgatagaaa cagaagccac tggagcacct caaaaacacc atcatacact
     4501 aaatcagtaa gttggcagca tcacccgacg cactttgcgc cgaataaata cctgtgacgg
     4561 aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg aagccctggg
     4621 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     4681 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     4741 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     4801 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     4861 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     4921 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattc
//
