LOCUS       sequence-288995-        7045 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 7045)
  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 7045)
  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-mut2_I
FEATURES             Location/Qualifiers
     source          1..7045
                     /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    3833..3954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3870..3889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3924..3943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3961..4416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4098..4119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4310..4329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4443..4547
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4549..4906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4549..4740
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4572..4592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4757..4892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4941..5735
                     /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             4941..5735
                     /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(4995..5014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5605..5624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5923..5942)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5967..6014
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6343..6931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6832..6851
                     /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 gaaaaacctc cgttataatt gcgaaaaccg caagcttact
     3001 ctccgatgtg gaggactatg aaagacgtta tctcctaagt ctggaagaag agcgacctgc
     3061 cctaatggat gacagaaagc acaaaatttg tagcatgtat gacaacttaa gggggaaatt
     3121 gcctggacaa gagaggccta gtgatgacca ctttgtacag atcatgtgta tccgaaaagg
     3181 gaagagaatg gttgcccgta ttcttccttt cctctccaca gagcaagcag ctgacattct
     3241 catgacaaca gccaggaacc tccctttcct tatcaagaag gatgcacaag atgaggtgct
     3301 gccatgctta ctgagtccct tctctctcct tctctatcat cttccatcag tgagtatcac
     3361 cagccttttg cgacagctaa tgaacctacc tcaaagtgca gctacaccag cactctccaa
     3421 tcctcacctc actgctgtgc tccagaacaa gtttggcctg tcactgctcc tcatcctcct
     3481 gagccgtggt gaagacctac agagttcaga ccctgctaca gaatcaacac aaaataatca
     3541 gtggacggag gtgatgttca tggcaacacg agaacttctg cggattcccc aagcagccct
     3601 ggccaagcca atctctatac ctacaaacct agtgtccctc ttttctcgct atgttgaccg
     3661 gcagaaactg aacttgctgg agacaaaact gcagctagtt caggggatac gataatctag
     3721 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3781 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3841 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3901 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3961 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4021 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4081 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4141 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4201 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4261 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4321 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4381 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4441 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4501 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4561 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4621 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4681 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4741 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4801 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4861 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4921 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4981 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5041 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5101 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5161 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5221 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5281 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5341 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5401 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5461 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5521 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5581 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5641 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5701 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5761 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5821 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5881 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5941 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6001 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6061 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6121 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6181 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6241 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6301 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6361 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6421 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6481 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6541 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6601 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6661 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6721 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6781 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6841 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6901 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6961 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7021 ggataaccgt attaccgcca tgcat
//
