LOCUS       sequence-289107-        5470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1_70-513.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5470)
  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 5470)
  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_70-513_J
FEATURES             Location/Qualifiers
     source          1..5470
                     /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    2258..2379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2295..2314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2349..2368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2386..2841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2523..2544)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2735..2754
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2868..2972
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2974..3331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2974..3165
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2997..3017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3182..3317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3366..4160
                     /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             3366..4160
                     /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(3420..3439)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4030..4049
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4348..4367)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4392..4439
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4768..5356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5257..5276
                     /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 agttcgataa tctagataac tgatcataat
     2161 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2221 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2281 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2341 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     2401 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2461 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2521 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2581 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2641 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2701 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2761 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2821 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2941 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3001 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3061 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3121 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3181 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3241 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3301 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3361 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3421 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3481 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3541 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3601 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3661 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3721 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3781 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3841 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3901 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3961 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4021 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4081 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4141 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4201 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4261 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4321 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4381 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4441 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4501 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4561 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4621 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4681 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4741 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4801 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4861 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4921 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4981 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5041 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5101 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5161 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5221 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5281 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5341 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5401 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5461 cgccatgcat
//
