LOCUS       sequence-289121-        4588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_517-770-R591AR595AK625AK626A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4588)
  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 4588)
  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-R591AR595AK625AK626A_J
FEATURES             Location/Qualifiers
     source          1..4588
                     /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        973..991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    992..1016
                     /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             1125..1169
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1207..1225)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1221..1268
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1498..2313)
                     /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(1498..2313)
                     /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     2221..2240
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2314..2405)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2421..3170)
                     /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(2431..2448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2671..2690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3366..4448)
                     /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     4410..4429
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4449..4526)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4572..4588,1..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 tatcgcaaaa gggaaggcaa tggttgcccg tattcttcct ttcctctcca cagagcaagc
      421 agctgacatt ctcatgacaa cagccaggaa cctccctttc cttatcgcag cagatgcaca
      481 agatgaggtg ctgccatgct tactgagtcc cttctctctc cttctctatc atcttccatc
      541 agtgagtatc accagccttt tgcgacagct aatgaaccta cctcaaagtg cagctacacc
      601 agcactctcc aatcctcacc tcactgctgt gctccagaac aagtttggcc tgtcactgct
      661 cctcatcctc ctgagccgtg gtgaagacct acagagttca gaccctgcta cagaatcaac
      721 acaaaataat cagtggacgg aggtgatgtt catggcaaca cgagaacttc tgcggattcc
      781 ccaagcagcc ctggccaagc caatctctat acctacaaac ctagtgtccc tcttttctcg
      841 ctatgttgac cggcagaaac tgaacttgct ggagacaaaa ctgcagctag ttcaggggat
      901 acgataagcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg tacacggccg
      961 cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccat
     1021 cttagtatat tagttaagta taagaaggag atatacatat ggcagatctc aattggatat
     1081 cggccggcca cgcgatcgct gacgtcggta ccctcgagtc tggtaaagaa accgctgctg
     1141 cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg
     1201 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1261 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     1321 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     1381 cgacaagctg acgaccgggt ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc
     1441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     1501 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1561 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1621 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1681 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1741 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     1801 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1861 ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1921 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     1981 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     2041 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     2101 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2161 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2221 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     2281 agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag
     2341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2401 acaaataggc atgcagcgct cttccgcttc ctcgctcact gactcgctac gctcggtcgt
     2461 tcgactgcgg cgagcggtgt cagctcactc aaaagcggta atacggttat ccacagaatc
     2521 aggggataaa gccggaaaga acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc
     2581 cgcaggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2641 gccagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2761 cttcgggaag cgtggcgctt tctcatagct cacgctgttg gtatctcagt tcggtgtagg
     2821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2941 cagccattgg taactgattt agaggacttt gtcttgaagt tatgcacctg ttaaggctaa
     3001 actgaaagaa cagattttgg tgagtgcggt cctccaaccc acttaccttg gttcaaagag
     3061 ttggtagctc agcgaacctt gagaaaacca ccgttggtag cggtggtttt tctttattta
     3121 tgagatgatg aatcaatcgg tctatcaagt caacgaacag ctattccgtt actctagatt
     3181 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     3241 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     3301 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     3361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     3481 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     3541 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     3601 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     3661 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     3721 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     3781 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     3841 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     3901 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     3961 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4021 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4081 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4141 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4201 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4261 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     4321 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     4381 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     4441 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     4501 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     4561 cattaggaaa ttaatacgac tcactata
//
