LOCUS       sequence-356591-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1_70-770.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  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 6290)
  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: pLNHA-C1-HsPatL1_70-770_J
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             665..730
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3078..3199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3115..3134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3169..3188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3206..3661)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3343..3364)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3555..3574
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3688..3792
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3794..4151
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3794..3985
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3817..3837)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4002..4137
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4186..4980
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4186..4980
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4240..4259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4850..4869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5168..5187)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5212..5259
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5588..6176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6077..6096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctaatg gagagagaga tgaaatggac ttgttgggtg accatgagga
      841 gaatctggca gaaaggctca gtaagatggt gattgaaaat gaactagaag atccagctat
      901 tatgagggca gtgcagacca ggccagtttt acaaccccaa ccaggaagtc tgaattccag
      961 tatctgggat ggatctgaag ttctgaggcg aatccgagga ccactgcttg ctcaggaaat
     1021 gcctacagtg tctgtattag aatatgcttt gcctcagagg cccccccagg gtccagaaga
     1081 tgatcgggac ctttctgaac gagcattacc aaggcggtca acttcaccta tcattggcag
     1141 tcctcctgtt agagctgtcc ccataggcac cccacctaaa cagatggctg tacccagctt
     1201 cacccaacag attctgtgtc cgaagcctgt ccatgttcgg cccccaatgc cacctcgtta
     1261 tcctgctccc tatggtgaga ggatgtctcc aaaccagctc tgcagtgtcc cgaactcttc
     1321 cctcctgggt cacccttttc ctcctagtgt tcctcctgtt ctcagccccc tccagagagc
     1381 acagcttctt ggaggagcac agctacagcc tggacggatg tctcccagcc agtttgcacg
     1441 ggtccctgga tttgttggta gtccacttgc tgccatgaat cccaagttgc tacaagggcg
     1501 agttgggcag atgcttcccc cagcaccagg cttccgtgcc ttctttagtg ctccaccctc
     1561 cgctacacca cctccacagc agcaccctcc tggcccagga ccccacctgc aaaacctaag
     1621 atctcaggcc ccaatgttta gaccggacac aactcacctc catccacagc accgtcgact
     1681 cttgcatcag agacagcaac agaatagaag tcagcatcgg aatctcaatg gtgcgggaga
     1741 tagaggaagt caccggagca gtcatcaaga tcatctccga aaggatccat atgccaatct
     1801 catgttgcag cgggaaaagg attgggtctc taaaatccag atgatgcaac tgcaaagcac
     1861 tgatccctac ctggatgatt tttattacca gaattacttt gaaaaactgg agaaactgtc
     1921 agctgctgaa gaaatacaag gtgatggccc taagaaggag cgcaccaagc ttatcacccc
     1981 tcaggtggcc aaactggagc acgcctataa gccagtgcaa tttgagggct ctttgggaaa
     2041 gcttaccgtt tctagtgtga ataatccccg aaaaatgatt gatgctgttg tgacatctcg
     2101 gagtgaggat gatgagacaa aagaaaaaca agttcgagac aagaggagaa aaacccttgt
     2161 tataattgag aaaacctaca gcttactcct tgatgtggag gactatgaaa gacgttatct
     2221 cctaagtctg gaagaagagc gacctgccct aatggatgac agaaagcaca aaatttgtag
     2281 catgtatgac aacttaaggg ggaaattgcc tggacaagag aggcctagtg atgaccactt
     2341 tgtacagatc atgtgtatcc gaaaagggaa gagaatggtt gcccgtattc ttcctttcct
     2401 ctccacagag caagcagctg acattctcat gacaacagcc aggaacctcc ctttccttat
     2461 caagaaggat gcacaagatg aggtgctgcc atgcttactg agtcccttct ctctccttct
     2521 ctatcatctt ccatcagtga gtatcaccag ccttttgcga cagctaatga acctacctca
     2581 aagtgcagct acaccagcac tctccaatcc tcacctcact gctgtgctcc agaacaagtt
     2641 tggcctgtca ctgctcctca tcctcctgag ccgtggtgaa gacctacaga gttcagaccc
     2701 tgctacagaa tcaacacaaa ataatcagtg gacggaggtg atgttcatgg caacacgaga
     2761 acttctgcgg attccccaag cagccctggc caagccaatc tctataccta caaacctagt
     2821 gtccctcttt tctcgctatg ttgaccggca gaaactgaac ttgctggaga caaaactgca
     2881 gctagttcag gggatacgat aatctagctc aagcttcgaa ttctgcagtc gacggtaccg
     2941 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3001 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3061 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3181 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3241 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3301 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3361 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3421 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3481 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3541 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3601 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3661 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3721 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3781 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3901 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3961 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4021 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4081 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4141 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4201 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4261 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4321 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4381 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4441 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4501 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4561 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4621 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4681 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4741 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4801 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4861 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4921 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4981 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5041 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5101 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5161 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5221 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5281 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5341 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5401 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5461 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5521 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5581 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5641 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5701 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5761 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5821 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5881 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5941 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6001 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6061 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6121 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6181 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6241 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
