LOCUS       sequence-289120-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_517-770-Mut3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4552)
  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 4552)
  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_517-770-Mut3_J
FEATURES             Location/Qualifiers
     source          1..4552
                     /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        937..955
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    956..980
                     /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             1089..1133
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1171..1189)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1185..1232
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1462..2277)
                     /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(1462..2277)
                     /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     2185..2204
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2278..2369)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2385..3134)
                     /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(2395..2412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2635..2654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3330..4412)
                     /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     4374..4393
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4413..4490)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4536..4552,1..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 tggaggacgg atctgggtct ggatctggga gaaagcacaa
      241 aatttgtagc atgtatgaca acttaagggg gaaattgcct ggacaagaga ggcctagtga
      301 tgaccacttt gtacagatca tgtgtatccg aaaagggaag agaatggttg cccgtattct
      361 tcctttcctc tccacagagc aagcagctga cattctcatg acaacagcca ggaacctccc
      421 tttccttatc aagaaggatg cacaagatga ggtgctgcca tgcttactga gtcccttctc
      481 tctccttctc tatcatcttc catcagtgag tatcaccagc cttttgcgac agctaatgaa
      541 cctacctcaa agtgcagcta caccagcact ctccaatcct cacctcactg ctgtgctcca
      601 gaacaagttt ggcctgtcac tgctcctcat cctcctgagc cgtggtgaag acctacagag
      661 ttcagaccct gctacagaat caacacaaaa taatcagtgg acggaggtga tgttcatggc
      721 aacacgagaa cttctgcgga ttccccaagc agccctggcc aagccaatct ctatacctac
      781 aaacctagtg tccctctttt ctcgctatgt tgaccggcag aaactgaact tgctggagac
      841 aaaactgcag ctagttcagg ggatacgata agcggccgca taatgcttaa gtcgaacaga
      901 aagtaatcgt attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat
      961 tgtgagcgga taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac
     1021 atatggcaga tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg
     1081 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     1141 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     1201 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
     1261 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     1321 tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact
     1381 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1441 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1501 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1561 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1621 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1681 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1741 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1801 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg
     1861 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1921 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     1981 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2041 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2101 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2161 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2221 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact
     2281 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2341 atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct
     2401 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc
     2461 ggtaatacgg ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag
     2521 caaagcaccg gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg
     2581 agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat
     2641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2761 gttggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2881 gacacgactt atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg
     2941 aagttatgca cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca
     3001 acccacttac cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg
     3061 gtagcggtgg tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga
     3121 acagctattc cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
     3181 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
     3241 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     3301 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3421 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     3481 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     3541 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     3601 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     3661 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     3721 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     3781 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     3841 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     3901 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     3961 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     4021 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     4081 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     4141 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     4201 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     4261 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     4321 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     4381 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     4441 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     4501 acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta ta
//
