LOCUS       sequence-289133-        7009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1-mut3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7009)
  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 7009)
  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: pT7-EGFP-C1-HsPatL1-mut3_J
FEATURES             Location/Qualifiers
     source          1..7009
                     /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             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3797..3918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3834..3853)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3888..3907
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3925..4380)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4062..4083)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4274..4293
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4407..4511
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4513..4870
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4513..4704
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4536..4556)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4721..4856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4905..5699
                     /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             4905..5699
                     /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(4959..4978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5569..5588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5887..5906)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5931..5978
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6307..6895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6796..6815
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctatgttccg ctacgagtct ttggaggatt gtcctctgga
     1441 tgaagatgaa gatgcatttc agggactggg agaagaagat gaagagattg atcaattcaa
     1501 tgatgataca tttgggtcag gtgcagttga tgatgattgg caggaagcac atgagcgcct
     1561 ggctgaattg gaagaaaagc taccagtggc agttaatgaa caaacaggca atggagagag
     1621 agatgaaatg gacttgttgg gtgaccatga ggagaatctg gcagaaaggc tcagtaagat
     1681 ggtgattgaa aatgaactag aagatccagc tattatgagg gcagtgcaga ccaggccagt
     1741 tttacaaccc caaccaggaa gtctgaattc cagtatctgg gatggatctg aagttctgag
     1801 gcgaatccga ggaccactgc ttgctcagga aatgcctaca gtgtctgtat tagaatatgc
     1861 tttgcctcag aggccccccc agggtccaga agatgatcgg gacctttctg aacgagcatt
     1921 accaaggcgg tcaacttcac ctatcattgg cagtcctcct gttagagctg tccccatagg
     1981 caccccacct aaacagatgg ctgtacccag cttcacccaa cagattctgt gtccgaagcc
     2041 tgtccatgtt cggcccccaa tgccacctcg ttatcctgct ccctatggtg agaggatgtc
     2101 tccaaaccag ctctgcagtg tcccgaactc ttccctcctg ggtcaccctt ttcctcctag
     2161 tgttcctcct gttctcagcc ccctccagag agcacagctt cttggaggag cacagctaca
     2221 gcctggacgg atgtctccca gccagtttgc acgggtccct ggatttgttg gtagtccact
     2281 tgctgccatg aatcccaagt tgctacaagg gcgagttggg cagatgcttc ccccagcacc
     2341 aggcttccgt gccttcttta gtgctccacc ctccgctaca ccacctccac agcagcaccc
     2401 tcctggccca ggaccccacc tgcaaaacct aagatctcag gccccaatgt ttagaccgga
     2461 cacaactcac ctccatccac agcaccgtcg actcttgcat cagagacagc aacagaatag
     2521 aagtcagcat cggaatctca atggtgcggg agatagagga agtcaccgga gcagtcatca
     2581 agatcatctc cgaaaggatc catatgccaa tctcatgttg cagcgggaaa aggattgggt
     2641 ctctaaaatc cagatgatgc aactgcaaag cactgatccc tacctggatg atttttatta
     2701 ccagaattac tttgaaaaac tggagaaact gtcagctgct gaagaaatac aaggtgatgg
     2761 ccctaagaag gagcgcacca agcttatcac ccctcaggtg gccaaactgg agcacgccta
     2821 taagccagtg caatttgagg gctctttggg aaagcttacc gtttctagtg tgaataatcc
     2881 ccgaaaaatg attgatgctg ttgtgacatc tcggagtgag gatgatgaga caaaagaaaa
     2941 acaagttcga gacaagagga gaaaaaccct tgttataatt gagaaaacct acagcttact
     3001 ccttgatgtg gaggacggat ctgggtctgg atctgggaga aagcacaaaa tttgtagcat
     3061 gtatgacaac ttaaggggga aattgcctgg acaagagagg cctagtgatg accactttgt
     3121 acagatcatg tgtatccgaa aagggaagag aatggttgcc cgtattcttc ctttcctctc
     3181 cacagagcaa gcagctgaca ttctcatgac aacagccagg aacctccctt tccttatcaa
     3241 gaaggatgca caagatgagg tgctgccatg cttactgagt cccttctctc tccttctcta
     3301 tcatcttcca tcagtgagta tcaccagcct tttgcgacag ctaatgaacc tacctcaaag
     3361 tgcagctaca ccagcactct ccaatcctca cctcactgct gtgctccaga acaagtttgg
     3421 cctgtcactg ctcctcatcc tcctgagccg tggtgaagac ctacagagtt cagaccctgc
     3481 tacagaatca acacaaaata atcagtggac ggaggtgatg ttcatggcaa cacgagaact
     3541 tctgcggatt ccccaagcag ccctggccaa gccaatctct atacctacaa acctagtgtc
     3601 cctcttttct cgctatgttg accggcagaa actgaacttg ctggagacaa aactgcagct
     3661 agttcagggg atacgataat ctagataact gatcataatc agccatacca catttgtaga
     3721 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3781 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3901 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3961 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4021 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4081 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4141 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4201 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4261 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4321 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4381 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4501 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4621 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4681 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4801 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4861 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4921 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4981 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5041 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5101 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5161 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5221 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5281 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5341 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5401 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     5461 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5521 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5581 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5641 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5701 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5761 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5821 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5881 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5941 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6001 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6061 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6121 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6181 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6241 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6421 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6961 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
