LOCUS       sequence-339294-        6497 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsPatL1.
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 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_I
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             605..670
                     /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             695..721
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3225..3346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3262..3281)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3316..3335
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3353..3808)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3490..3511)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3702..3721
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3835..3939
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3941..4298
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3941..4132
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3964..3984)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4149..4284
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4333..5127
                     /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             4333..5127
                     /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(4387..4406)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4997..5016
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5315..5334)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5359..5406
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5735..6323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6224..6243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      661 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      721 tcctccatct cgagctatgt tccgctacga gtctttggag gattgtcctc tggatgaaga
      781 tgaagatgca tttcagggac tgggagaaga agatgaagag attgatcaat tcaatgatga
      841 tacatttggg tcaggtgcag ttgatgatga ttggcaggaa gcacatgagc gcctggctga
      901 attggaagaa aagctaccag tggcagttaa tgaacaaaca ggcaatggag agagagatga
      961 aatggacttg ttgggtgacc atgaggagaa tctggcagaa aggctcagta agatggtgat
     1021 tgaaaatgaa ctagaagatc cagctattat gagggcagtg cagaccaggc cagttttaca
     1081 accccaacca ggaagtctga attccagtat ctgggatgga tctgaagttc tgaggcgaat
     1141 ccgaggacca ctgcttgctc aggaaatgcc tacagtgtct gtattagaat atgctttgcc
     1201 tcagaggccc ccccagggtc cagaagatga tcgggacctt tctgaacgag cattaccaag
     1261 gcggtcaact tcacctatca ttggcagtcc tcctgttaga gctgtcccca taggcacccc
     1321 acctaaacag atggctgtac ccagcttcac ccaacagatt ctgtgtccga agcctgtcca
     1381 tgttcggccc ccaatgccac ctcgttatcc tgctccctat ggtgagagga tgtctccaaa
     1441 ccagctctgc agtgtcccga actcttccct cctgggtcac ccttttcctc ctagtgttcc
     1501 tcctgttctc agccccctcc agagagcaca gcttcttgga ggagcacagc tacagcctgg
     1561 acggatgtct cccagccagt ttgcacgggt ccctggattt gttggtagtc cacttgctgc
     1621 catgaatccc aagttgctac aagggcgagt tgggcagatg cttcccccag caccaggctt
     1681 ccgtgccttc tttagtgctc caccctccgc tacaccacct ccacagcagc accctcctgg
     1741 cccaggaccc cacctgcaaa acctaagatc tcaggcccca atgtttagac cggacacaac
     1801 tcacctccat ccacagcacc gtcgactctt gcatcagaga cagcaacaga atagaagtca
     1861 gcatcggaat ctcaatggtg cgggagatag aggaagtcac cggagcagtc atcaagatca
     1921 tctccgaaag gatccatatg ccaatctcat gttgcagcgg gaaaaggatt gggtctctaa
     1981 aatccagatg atgcaactgc aaagcactga tccctacctg gatgattttt attaccagaa
     2041 ttactttgaa aaactggaga aactgtcagc tgctgaagaa atacaaggtg atggccctaa
     2101 gaaggagcgc accaagctta tcacccctca ggtggccaaa ctggagcacg cctataagcc
     2161 agtgcaattt gagggctctt tgggaaagct taccgtttct agtgtgaata atccccgaaa
     2221 aatgattgat gctgttgtga catctcggag tgaggatgat gagacaaaag aaaaacaagt
     2281 tcgagacaag aggagaaaaa cccttgttat aattgagaaa acctacagct tactccttga
     2341 tgtggaggac tatgaaagac gttatctcct aagtctggaa gaagagcgac ctgccctaat
     2401 ggatgacaga aagcacaaaa tttgtagcat gtatgacaac ttaaggggga aattgcctgg
     2461 acaagagagg cctagtgatg accactttgt acagatcatg tgtatccgaa aagggaagag
     2521 aatggttgcc cgtattcttc ctttcctctc cacagagcaa gcagctgaca ttctcatgac
     2581 aacagccagg aacctccctt tccttatcaa gaaggatgca caagatgagg tgctgccatg
     2641 cttactgagt cccttctctc tccttctcta tcatcttcca tcagtgagta tcaccagcct
     2701 tttgcgacag ctaatgaacc tacctcaaag tgcagctaca ccagcactct ccaatcctca
     2761 cctcactgct gtgctccaga acaagtttgg cctgtcactg ctcctcatcc tcctgagccg
     2821 tggtgaagac ctacagagtt cagaccctgc tacagaatca acacaaaata atcagtggac
     2881 ggaggtgatg ttcatggcaa cacgagaact tctgcggatt ccccaagcag ccctggccaa
     2941 gccaatctct atacctacaa acctagtgtc cctcttttct cgctatgttg accggcagaa
     3001 actgaacttg ctggagacaa aactgcagct agttcagggg atacgataat ctagctcaag
     3061 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     3121 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3181 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3241 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3301 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3361 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3421 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3481 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3541 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3601 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3661 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3721 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3781 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3961 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4141 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4201 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4261 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4321 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4381 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4441 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4501 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4561 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4621 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4681 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4741 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4801 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4861 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4921 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4981 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5041 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5101 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5161 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5221 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5281 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5341 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5401 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5461 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5521 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5581 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5641 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5881 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6421 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     6481 ccatatatgg agttccg
//
