LOCUS       sequence-291015-        4958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of Hs4EHP_52-234opt-Q159SM161DR202E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4958)
  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 4958)
  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-NpH-Hs4EHP_52-234opt-Q159SM161DR202E_AE
FEATURES             Location/Qualifiers
     source          1..4958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2294..2313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2402..2421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2402..2420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2421..2445
                     /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             2460..2482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2501..2518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2540..2563
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3190..3208)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3204..3251
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3426..3431,3432..3448,3449..3454))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3566..4111
                     /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     4012..4031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4228..4245
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4637..4739
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4740..4958,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4806..4825)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
      541 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
      601 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
      661 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
      721 gagcctgata aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg
      841 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg
      901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
      961 cggaaggagc taccggacag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tgactctcag tcatagtatc gtggtatcac cggttggttc cactctctgt
     1081 tgcgggcaac ttcagcagca cgtaggggac ttccgcgttt ccagacttta cgaaacacgg
     1141 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1201 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1261 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1321 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1381 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1441 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1501 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1561 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccagtga
     1621 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1681 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1741 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1801 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1861 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1921 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1981 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     2041 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     2101 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2161 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2221 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2281 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2341 tgatgccggc cacgatgcgt ccggcgtaga ggatcagact aagatctcga tcccgcgaaa
     2401 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccact agtaataatt
     2461 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
     2521 cagcggtacc ggcagcggcc tggaagttct gttccagggg ccccatatgc tcgaggcaga
     2581 acatccgctg cagtataact ataccttttg gtatagccgt cgtacaccgg gtcgtccgac
     2641 cagcagccag agctatgaac agaacattaa acaaattggc acctttgcaa gcgtggaaca
     2701 gttttggcgt ttttatagcc atatggttcg tccgggtgat ctgacaggtc atagcgattt
     2761 tcacctgttt aaagaaggca ttaaaccgat gtgggaagat gatgcaaaca aaaatggtgg
     2821 caaatggatt attcgtctgc gtaaaggtct ggcaagccgt tgttgggaaa atctgattct
     2881 ggcaatgctg ggtgaatcgt ttgacgttgg tgaagaaatt tgtggtgcag ttgttagcgt
     2941 tcgttttcaa gaagatatta tcagcatctg gaataaaacc gcaagcgatc aggcaaccac
     3001 cgcacgtatt cgtgataccc tgcgtgaggt tctgaatctg cctccgaata ccattatgga
     3061 atacaaaacc cataccgata gcatcaaaat gcctggtcgt ctgggtccgc agcgtctgct
     3121 gttttgagct agccctagga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     3181 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     3241 ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc ccggcagtac
     3301 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
     3361 cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact
     3421 accgcattaa agcttatcga tgataagctg tcaaacatga gaattacaac ttatatcgta
     3481 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc aagaaatatt
     3541 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     3601 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     3661 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     3721 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     3781 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     3841 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     3901 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     3961 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     4021 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     4081 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     4141 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     4201 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     4261 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     4321 cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc cctagcgctg
     4381 atgtccggcg gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca
     4441 gctgatagaa acagaagcca ctggagcacc tcaaaaacac catcatacac taaatcagta
     4501 agttggcagc atcacccgac gcactttgcg ccgaataaat acctgtgacg gaagatcact
     4561 tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt
     4621 ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     4681 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     4741 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     4801 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     4861 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     4921 acattcttgc ccgcctgatg aatgctcatc cggaattc
//
