LOCUS       sequence-289117-        4555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_517-767-del664-673.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4555)
  AUTHORS   Braun JE, Tritschler F, Haas G, Igreja C, Truffault V, Weichenrieder
            O, Izaurralde E
  TITLE     The C-terminal alpha-alpha superhelix of Pat is required for mRNA 
            decapping in metazoa.
  JOURNAL   EMBO J. 2010 Jul 21. 29(14):2368-80.
  PUBMED    20543818
REFERENCE   2  (bases 1 to 4555)
  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"; journalName: "EMBO J. 
            2010 Jul 21. 29(14):2368-80."
COMMENT     Sequence Label: pRSFDuetP-HsPatL1_517-767del664-673_J
FEATURES             Location/Qualifiers
     source          1..4555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..133
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        940..958
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    959..983
                     /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)."
     CDS             1092..1136
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1174..1192)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1188..1235
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1465..2280)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1465..2280)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2188..2207
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2281..2372)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2388..3137)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2398..2415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2638..2657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3333..4415)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4377..4396
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4416..4493)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4539..4555,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agcgcgggtc tggaagttct
      121 gttccagggg ccccaggatc cggacaagag gagaaaaacc cttgttataa ttgagaaaac
      181 ctacagctta ctccttgatg tggaggacta tgaaagacgt tatctcctaa gtctggaaga
      241 agagcgacct gccctaatgg atgacagaaa gcacaaaatt tgtagcatgt atgacaactt
      301 aagggggaaa ttgcctggac aagagaggcc tagtgatgac cactttgtac agatcatgtg
      361 tatccgaaaa gggaagagaa tggttgcccg tattcttcct ttcctctcca cagagcaagc
      421 agctgacatt ctcatgacaa cagccaggaa cctccctttc cttatcaaga aggatgcaca
      481 agatgaggtg ctgccatgct tactgagtcc cttctctctc cttctctatc atcttccatc
      541 agtgagtatc accagccttt tgcgacagct aatgaaccta cctgggtctc ctcacctcac
      601 tgctgtgctc cagaacaagt ttggcctgtc actgctcctc atcctcctga gccgtggtga
      661 agacctacag agttcagacc ctgctacaga atcaacacaa aataatcagt ggacggaggt
      721 gatgttcatg gcaacacgag aacttctgcg gattccccaa gcagccctgg ccaagccaat
      781 ctctatacct acaaacctag tgtccctctt ttctcgctat gttgaccggc agaaactgaa
      841 cttgctggag acaaaactgc agctagttca gtaagcggcc gcataatgct taagtcgaac
      901 agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg
      961 aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata
     1021 tacatatggc agatctcaat tggatatcgg ccggccacgc gatcgctgac gtcggtaccc
     1081 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     1141 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     1201 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     1261 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     1321 ctatttaacg accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc
     1381 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1441 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     1501 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     1561 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     1621 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     1681 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     1741 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     1801 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa
     1861 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     1921 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     1981 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     2041 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2101 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     2161 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     2221 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     2281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2341 catatttgaa tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc
     2401 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa
     2461 agcggtaata cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa
     2521 aagcaaagca ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg
     2581 acgagcatca caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa
     2641 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2701 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2761 gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2821 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2881 taagacacga cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc
     2941 ttgaagttat gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct
     3001 ccaacccact taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg
     3061 ttggtagcgg tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa
     3121 cgaacagcta ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     3181 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     3241 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3301 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3361 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3421 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3481 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3541 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     3601 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     3661 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     3721 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     3781 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     3841 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     3901 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     3961 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4021 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4081 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4141 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4201 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4261 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4321 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4381 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4441 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     4501 tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca ctata
//
