LOCUS       sequence-289115-        6461 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsPatL1-mut3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6461)
  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 6461)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-mut3_J
FEATURES             Location/Qualifiers
     source          1..6461
                     /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    3249..3370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3286..3305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3340..3359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3377..3832)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3514..3535)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3726..3745
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3859..3963
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3965..4322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3965..4156
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3988..4008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4173..4308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4357..5151
                     /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             4357..5151
                     /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(4411..4430)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5021..5040
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5339..5358)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5383..5430
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5759..6347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6248..6267
                     /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 cccttgttat aattgagaaa acctacagct tactccttga
     2401 tgtggaggac ggatctgggt ctggatctgg gagaaagcac aaaatttgta gcatgtatga
     2461 caacttaagg gggaaattgc ctggacaaga gaggcctagt gatgaccact ttgtacagat
     2521 catgtgtatc cgaaaaggga agagaatggt tgcccgtatt cttcctttcc tctccacaga
     2581 gcaagcagct gacattctca tgacaacagc caggaacctc cctttcctta tcaagaagga
     2641 tgcacaagat gaggtgctgc catgcttact gagtcccttc tctctccttc tctatcatct
     2701 tccatcagtg agtatcacca gccttttgcg acagctaatg aacctacctc aaagtgcagc
     2761 tacaccagca ctctccaatc ctcacctcac tgctgtgctc cagaacaagt ttggcctgtc
     2821 actgctcctc atcctcctga gccgtggtga agacctacag agttcagacc ctgctacaga
     2881 atcaacacaa aataatcagt ggacggaggt gatgttcatg gcaacacgag aacttctgcg
     2941 gattccccaa gcagccctgg ccaagccaat ctctatacct acaaacctag tgtccctctt
     3001 ttctcgctat gttgaccggc agaaactgaa cttgctggag acaaaactgc agctagttca
     3061 ggggatacga taatctagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
     3121 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     3181 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3241 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3301 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3361 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3421 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3481 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3541 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3601 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3661 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3721 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3781 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3841 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3901 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3961 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4141 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4201 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4261 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4321 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4381 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4441 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4501 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4561 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4621 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4681 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4741 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4801 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4861 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4921 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4981 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5041 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5101 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5161 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5221 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5281 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5341 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5401 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5461 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5521 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5581 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5641 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5701 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5761 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5821 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5881 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5941 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6001 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6061 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6121 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6181 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6241 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6301 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6361 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6421 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
