LOCUS       sequence-289128-        6487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1_70-770del399-516.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6487)
  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 6487)
  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_70-770del399-516_J
FEATURES             Location/Qualifiers
     source          1..6487
                     /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    3275..3396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3312..3331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3366..3385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3403..3858)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3540..3561)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3752..3771
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3885..3989
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3991..4348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3991..4182
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4014..4034)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4199..4334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4383..5177
                     /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             4383..5177
                     /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(4437..4456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5047..5066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5365..5384)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5409..5456
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5785..6373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6274..6293
                     /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 ctaatggaga gagagatgaa atggacttgt tgggtgacca
     1441 tgaggagaat ctggcagaaa ggctcagtaa gatggtgatt gaaaatgaac tagaagatcc
     1501 agctattatg agggcagtgc agaccaggcc agttttacaa ccccaaccag gaagtctgaa
     1561 ttccagtatc tgggatggat ctgaagttct gaggcgaatc cgaggaccac tgcttgctca
     1621 ggaaatgcct acagtgtctg tattagaata tgctttgcct cagaggcccc cccagggtcc
     1681 agaagatgat cgggaccttt ctgaacgagc attaccaagg cggtcaactt cacctatcat
     1741 tggcagtcct cctgttagag ctgtccccat aggcacccca cctaaacaga tggctgtacc
     1801 cagcttcacc caacagattc tgtgtccgaa gcctgtccat gttcggcccc caatgccacc
     1861 tcgttatcct gctccctatg gtgagaggat gtctccaaac cagctctgca gtgtcccgaa
     1921 ctcttccctc ctgggtcacc cttttcctcc tagtgttcct cctgttctca gccccctcca
     1981 gagagcacag cttcttggag gagcacagct acagcctgga cggatgtctc ccagccagtt
     2041 tgcacgggtc cctggatttg ttggtagtcc acttgctgcc atgaatccca agttgctaca
     2101 agggcgagtt gggcagatgc ttcccccagc accaggcttc cgtgccttct ttagtgctcc
     2161 accctccgct acaccacctc cacagcagca ccctcctggc ccaggacccc acctgcaaaa
     2221 cctaagatct caggccccaa tgtttagacc ggacacaact cacctccatc cacagcaccg
     2281 tcgactcttg catcagagac agcaacagaa tagaagtcag catcggaatc tcaatggtgc
     2341 gggagataga ggaagtcacc ggagcagtca tcaagatcat ctccgaaagg atgacaagag
     2401 gagaaaaacc cttgttataa ttgagaaaac ctacagctta ctccttgatg tggaggacta
     2461 tgaaagacgt tatctcctaa gtctggaaga agagcgacct gccctaatgg atgacagaaa
     2521 gcacaaaatt tgtagcatgt atgacaactt aagggggaaa ttgcctggac aagagaggcc
     2581 tagtgatgac cactttgtac agatcatgtg tatccgaaaa gggaagagaa tggttgcccg
     2641 tattcttcct ttcctctcca cagagcaagc agctgacatt ctcatgacaa cagccaggaa
     2701 cctccctttc cttatcaaga aggatgcaca agatgaggtg ctgccatgct tactgagtcc
     2761 cttctctctc cttctctatc atcttccatc agtgagtatc accagccttt tgcgacagct
     2821 aatgaaccta cctcaaagtg cagctacacc agcactctcc aatcctcacc tcactgctgt
     2881 gctccagaac aagtttggcc tgtcactgct cctcatcctc ctgagccgtg gtgaagacct
     2941 acagagttca gaccctgcta cagaatcaac acaaaataat cagtggacgg aggtgatgtt
     3001 catggcaaca cgagaacttc tgcggattcc ccaagcagcc ctggccaagc caatctctat
     3061 acctacaaac ctagtgtccc tcttttctcg ctatgttgac cggcagaaac tgaacttgct
     3121 ggagacaaaa ctgcagctag ttcaggggat acgataatct agataactga tcataatcag
     3181 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3361 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3421 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3481 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3541 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3601 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3661 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3721 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3781 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3841 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3901 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3961 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4021 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4081 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4141 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4201 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4261 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4321 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4381 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4441 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4501 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4561 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4621 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4681 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4741 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4801 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4861 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4921 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4981 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5041 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5101 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5161 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5221 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5281 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5341 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5401 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5461 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5521 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5581 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5641 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5701 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6421 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6481 catgcat
//
