LOCUS       sequence-288992-        6497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1-mut2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  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 6497)
  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-mut2_I
FEATURES             Location/Qualifiers
     source          1..6497
                     /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    3285..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3322..3341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3376..3395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3413..3868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3571)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3762..3781
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3895..3999
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4001..4358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4001..4192
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4024..4044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4209..4344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4393..5187
                     /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             4393..5187
                     /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(4447..4466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5057..5076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5375..5394)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5419..5466
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6284..6303
                     /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 cgagctatgt tccgctacga gtctttggag gattgtcctc tggatgaaga
      841 tgaagatgca tttcagggac tgggagaaga agatgaagag attgatcaat tcaatgatga
      901 tacatttggg tcaggtgcag ttgatgatga ttggcaggaa gcacatgagc gcctggctga
      961 attggaagaa aagctaccag tggcagttaa tgaacaaaca ggcaatggag agagagatga
     1021 aatggacttg ttgggtgacc atgaggagaa tctggcagaa aggctcagta agatggtgat
     1081 tgaaaatgaa ctagaagatc cagctattat gagggcagtg cagaccaggc cagttttaca
     1141 accccaacca ggaagtctga attccagtat ctgggatgga tctgaagttc tgaggcgaat
     1201 ccgaggacca ctgcttgctc aggaaatgcc tacagtgtct gtattagaat atgctttgcc
     1261 tcagaggccc ccccagggtc cagaagatga tcgggacctt tctgaacgag cattaccaag
     1321 gcggtcaact tcacctatca ttggcagtcc tcctgttaga gctgtcccca taggcacccc
     1381 acctaaacag atggctgtac ccagcttcac ccaacagatt ctgtgtccga agcctgtcca
     1441 tgttcggccc ccaatgccac ctcgttatcc tgctccctat ggtgagagga tgtctccaaa
     1501 ccagctctgc agtgtcccga actcttccct cctgggtcac ccttttcctc ctagtgttcc
     1561 tcctgttctc agccccctcc agagagcaca gcttcttgga ggagcacagc tacagcctgg
     1621 acggatgtct cccagccagt ttgcacgggt ccctggattt gttggtagtc cacttgctgc
     1681 catgaatccc aagttgctac aagggcgagt tgggcagatg cttcccccag caccaggctt
     1741 ccgtgccttc tttagtgctc caccctccgc tacaccacct ccacagcagc accctcctgg
     1801 cccaggaccc cacctgcaaa acctaagatc tcaggcccca atgtttagac cggacacaac
     1861 tcacctccat ccacagcacc gtcgactctt gcatcagaga cagcaacaga atagaagtca
     1921 gcatcggaat ctcaatggtg cgggagatag aggaagtcac cggagcagtc atcaagatca
     1981 tctccgaaag gatccatatg ccaatctcat gttgcagcgg gaaaaggatt gggtctctaa
     2041 aatccagatg atgcaactgc aaagcactga tccctacctg gatgattttt attaccagaa
     2101 ttactttgaa aaactggaga aactgtcagc tgctgaagaa atacaaggtg atggccctaa
     2161 gaaggagcgc accaagctta tcacccctca ggtggccaaa ctggagcacg cctataagcc
     2221 agtgcaattt gagggctctt tgggaaagct taccgtttct agtgtgaata atccccgaaa
     2281 aatgattgat gctgttgtga catctcggag tgaggatgat gagacaaaag aaaaacaagt
     2341 tcgagacaag aggagaaaaa cctccgttat aattgcgaaa accgcaagct tactctccga
     2401 tgtggaggac tatgaaagac gttatctcct aagtctggaa gaagagcgac ctgccctaat
     2461 ggatgacaga aagcacaaaa tttgtagcat gtatgacaac ttaaggggga aattgcctgg
     2521 acaagagagg cctagtgatg accactttgt acagatcatg tgtatccgaa aagggaagag
     2581 aatggttgcc cgtattcttc ctttcctctc cacagagcaa gcagctgaca ttctcatgac
     2641 aacagccagg aacctccctt tccttatcaa gaaggatgca caagatgagg tgctgccatg
     2701 cttactgagt cccttctctc tccttctcta tcatcttcca tcagtgagta tcaccagcct
     2761 tttgcgacag ctaatgaacc tacctcaaag tgcagctaca ccagcactct ccaatcctca
     2821 cctcactgct gtgctccaga acaagtttgg cctgtcactg ctcctcatcc tcctgagccg
     2881 tggtgaagac ctacagagtt cagaccctgc tacagaatca acacaaaata atcagtggac
     2941 ggaggtgatg ttcatggcaa cacgagaact tctgcggatt ccccaagcag ccctggccaa
     3001 gccaatctct atacctacaa acctagtgtc cctcttttct cgctatgttg accggcagaa
     3061 actgaacttg ctggagacaa aactgcagct agttcagggg atacgataat ctagctcaag
     3121 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     3181 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3241 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3421 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3481 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3541 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3601 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3661 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3721 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3781 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3841 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3901 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3961 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4021 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4201 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4261 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4321 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4381 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4561 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4921 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5221 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5281 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5341 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5401 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5461 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5521 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5581 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5641 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5701 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtattaccgc catgcat
//
