LOCUS       sequence-357821-        4527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of Hs4EHP_52-234opt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4527)
  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 4527)
  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_AE
FEATURES             Location/Qualifiers
     source          1..4527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             653..673
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             701..718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(799..817)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      813..860
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1035..1040,1041..1057,1058..1063))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      1175..1720
                     /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     1621..1640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1837..1854
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2246..2348
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2349..3008
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2415..2434)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4420..4439
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
       61 tgtttaactt taagaaggag atatacatat gctcgaggca gaacatccgc tgcagtataa
      121 ctataccttt tggtatagcc gtcgtacacc gggtcgtccg accagcagcc agagctatga
      181 acagaacatt aaacaaattg gcacctttgc aagcgtggaa cagttttggc gtttttatag
      241 ccatatggtt cgtccgggtg atctgacagg tcatagcgat tttcacctgt ttaaagaagg
      301 cattaaaccg atgtgggaag atgatgcaaa caaaaatggt ggcaaatgga ttattcgtct
      361 gcgtaaaggt ctggcaagcc gttgttggga aaatctgatt ctggcaatgc tgggtgaaca
      421 gtttatggtt ggtgaagaaa tttgtggtgc agttgttagc gttcgttttc aagaagatat
      481 tatcagcatc tggaataaaa ccgcaagcga tcaggcaacc accgcacgta ttcgtgatac
      541 cctgcgtcgt gttctgaatc tgcctccgaa taccattatg gaatacaaaa cccataccga
      601 tagcatcaaa atgcctggtc gtctgggtcc gcagcgtctg ctgtttggat ccgaaaacct
      661 ttacttccag ggcagcggcg ccatgggcag cggcagcggc caccaccacc accaccacgg
      721 tacctaatag tctagagcta gccctaggag atccggctgc taacaaagcc cgaaaggaag
      781 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      841 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
      901 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
      961 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     1021 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattacaact
     1081 tatatcgtat ggggctgact tcaggtgcta catttgaaga gataaattgc actgaaatca
     1141 agaaatattt tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
     1201 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
     1261 ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc
     1321 tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca
     1381 gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta
     1441 ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag
     1501 cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag
     1561 cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg
     1621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca
     1681 gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct
     1741 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
     1801 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     1861 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
     1921 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccc
     1981 ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcag tagctgaaca
     2041 ggagggacag ctgatagaaa cagaagccac tggagcacct caaaaacacc atcatacact
     2101 aaatcagtaa gttggcagca tcacccgacg cactttgcgc cgaataaata cctgtgacgg
     2161 aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg aagccctggg
     2221 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     2281 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     2341 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     2401 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     2461 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     2521 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
     2581 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     2641 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     2701 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2761 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2821 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2881 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     2941 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     3001 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     3061 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc cggtcgtcgg
     3121 ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac
     3181 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
     3241 ccccgtggag gtaataattg acgatatgat catttattct gcctcccaga gcctgataaa
     3301 aacggttagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     3361 gcgatgcaga tccggaacat aatggtgcag ggcgcttgtt tcggcgtggg tatggtggca
     3421 ggccccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
     3481 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
     3541 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
     3601 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
     3661 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
     3721 cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc
     3781 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
     3841 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
     3901 cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg
     3961 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     4021 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     4081 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     4141 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     4201 gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca
     4261 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     4321 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     4381 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     4441 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     4501 cagactaaga tctcgatccc gcgaaat
//
