LOCUS       sequence-339295-        7045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1.
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_I
FEATURES             Location/Qualifiers
     source          1..7045
                     /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             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /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(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3773..3894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3810..3829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3864..3883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3901..4356)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4038..4059)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4250..4269
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4383..4487
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4489..4846
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4489..4680
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4512..4532)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4697..4832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4881..5675
                     /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             4881..5675
                     /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(4935..4954)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5545..5564
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5863..5882)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5907..5954
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6283..6871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6772..6791
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag ctatgttccg ctacgagtct ttggaggatt gtcctctgga
     1381 tgaagatgaa gatgcatttc agggactggg agaagaagat gaagagattg atcaattcaa
     1441 tgatgataca tttgggtcag gtgcagttga tgatgattgg caggaagcac atgagcgcct
     1501 ggctgaattg gaagaaaagc taccagtggc agttaatgaa caaacaggca atggagagag
     1561 agatgaaatg gacttgttgg gtgaccatga ggagaatctg gcagaaaggc tcagtaagat
     1621 ggtgattgaa aatgaactag aagatccagc tattatgagg gcagtgcaga ccaggccagt
     1681 tttacaaccc caaccaggaa gtctgaattc cagtatctgg gatggatctg aagttctgag
     1741 gcgaatccga ggaccactgc ttgctcagga aatgcctaca gtgtctgtat tagaatatgc
     1801 tttgcctcag aggccccccc agggtccaga agatgatcgg gacctttctg aacgagcatt
     1861 accaaggcgg tcaacttcac ctatcattgg cagtcctcct gttagagctg tccccatagg
     1921 caccccacct aaacagatgg ctgtacccag cttcacccaa cagattctgt gtccgaagcc
     1981 tgtccatgtt cggcccccaa tgccacctcg ttatcctgct ccctatggtg agaggatgtc
     2041 tccaaaccag ctctgcagtg tcccgaactc ttccctcctg ggtcaccctt ttcctcctag
     2101 tgttcctcct gttctcagcc ccctccagag agcacagctt cttggaggag cacagctaca
     2161 gcctggacgg atgtctccca gccagtttgc acgggtccct ggatttgttg gtagtccact
     2221 tgctgccatg aatcccaagt tgctacaagg gcgagttggg cagatgcttc ccccagcacc
     2281 aggcttccgt gccttcttta gtgctccacc ctccgctaca ccacctccac agcagcaccc
     2341 tcctggccca ggaccccacc tgcaaaacct aagatctcag gccccaatgt ttagaccgga
     2401 cacaactcac ctccatccac agcaccgtcg actcttgcat cagagacagc aacagaatag
     2461 aagtcagcat cggaatctca atggtgcggg agatagagga agtcaccgga gcagtcatca
     2521 agatcatctc cgaaaggatc catatgccaa tctcatgttg cagcgggaaa aggattgggt
     2581 ctctaaaatc cagatgatgc aactgcaaag cactgatccc tacctggatg atttttatta
     2641 ccagaattac tttgaaaaac tggagaaact gtcagctgct gaagaaatac aaggtgatgg
     2701 ccctaagaag gagcgcacca agcttatcac ccctcaggtg gccaaactgg agcacgccta
     2761 taagccagtg caatttgagg gctctttggg aaagcttacc gtttctagtg tgaataatcc
     2821 ccgaaaaatg attgatgctg ttgtgacatc tcggagtgag gatgatgaga caaaagaaaa
     2881 acaagttcga gacaagagga gaaaaaccct tgttataatt gagaaaacct acagcttact
     2941 ccttgatgtg gaggactatg aaagacgtta tctcctaagt ctggaagaag agcgacctgc
     3001 cctaatggat gacagaaagc acaaaatttg tagcatgtat gacaacttaa gggggaaatt
     3061 gcctggacaa gagaggccta gtgatgacca ctttgtacag atcatgtgta tccgaaaagg
     3121 gaagagaatg gttgcccgta ttcttccttt cctctccaca gagcaagcag ctgacattct
     3181 catgacaaca gccaggaacc tccctttcct tatcaagaag gatgcacaag atgaggtgct
     3241 gccatgctta ctgagtccct tctctctcct tctctatcat cttccatcag tgagtatcac
     3301 cagccttttg cgacagctaa tgaacctacc tcaaagtgca gctacaccag cactctccaa
     3361 tcctcacctc actgctgtgc tccagaacaa gtttggcctg tcactgctcc tcatcctcct
     3421 gagccgtggt gaagacctac agagttcaga ccctgctaca gaatcaacac aaaataatca
     3481 gtggacggag gtgatgttca tggcaacacg agaacttctg cggattcccc aagcagccct
     3541 ggccaagcca atctctatac ctacaaacct agtgtccctc ttttctcgct atgttgaccg
     3601 gcagaaactg aacttgctgg agacaaaact gcagctagtt caggggatac gataatctag
     3661 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3721 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3901 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3961 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4021 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4081 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4141 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4201 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4261 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4321 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4381 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4441 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4501 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4561 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4621 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4681 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4741 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4801 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4861 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4921 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4981 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5041 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5101 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5161 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5221 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5281 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5341 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5401 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5461 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5521 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5581 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5641 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5701 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5761 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5821 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5881 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5941 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6001 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6061 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6121 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6181 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6241 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6301 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6361 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6421 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6481 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6541 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6601 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6661 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6721 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6781 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6841 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6901 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6961 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     7021 ttcatagccc atatatggag ttccg
//
