LOCUS       sequence-289130-        5851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1_399-770.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5851)
  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 5851)
  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_399-770_J
FEATURES             Location/Qualifiers
     source          1..5851
                     /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    2639..2760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2676..2695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2730..2749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2767..3222)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2904..2925)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3116..3135
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3249..3353
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3355..3712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3355..3546
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3378..3398)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3563..3698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3747..4541
                     /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             3747..4541
                     /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(3801..3820)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4411..4430
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4729..4748)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4773..4820
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5149..5737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5638..5657
                     /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 ctgatccata tgccaatctc atgttgcagc gggaaaagga
     1441 ttgggtctct aaaatccaga tgatgcaact gcaaagcact gatccctacc tggatgattt
     1501 ttattaccag aattactttg aaaaactgga gaaactgtca gctgctgaag aaatacaagg
     1561 tgatggccct aagaaggagc gcaccaagct tatcacccct caggtggcca aactggagca
     1621 cgcctataag ccagtgcaat ttgagggctc tttgggaaag cttaccgttt ctagtgtgaa
     1681 taatccccga aaaatgattg atgctgttgt gacatctcgg agtgaggatg atgagacaaa
     1741 agaaaaacaa gttcgagaca agaggagaaa aacccttgtt ataattgaga aaacctacag
     1801 cttactcctt gatgtggagg actatgaaag acgttatctc ctaagtctgg aagaagagcg
     1861 acctgcccta atggatgaca gaaagcacaa aatttgtagc atgtatgaca acttaagggg
     1921 gaaattgcct ggacaagaga ggcctagtga tgaccacttt gtacagatca tgtgtatccg
     1981 aaaagggaag agaatggttg cccgtattct tcctttcctc tccacagagc aagcagctga
     2041 cattctcatg acaacagcca ggaacctccc tttccttatc aagaaggatg cacaagatga
     2101 ggtgctgcca tgcttactga gtcccttctc tctccttctc tatcatcttc catcagtgag
     2161 tatcaccagc cttttgcgac agctaatgaa cctacctcaa agtgcagcta caccagcact
     2221 ctccaatcct cacctcactg ctgtgctcca gaacaagttt ggcctgtcac tgctcctcat
     2281 cctcctgagc cgtggtgaag acctacagag ttcagaccct gctacagaat caacacaaaa
     2341 taatcagtgg acggaggtga tgttcatggc aacacgagaa cttctgcgga ttccccaagc
     2401 agccctggcc aagccaatct ctatacctac aaacctagtg tccctctttt ctcgctatgt
     2461 tgaccggcag aaactgaact tgctggagac aaaactgcag ctagttcagg ggatacgata
     2521 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2581 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2641 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2701 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2761 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2821 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2881 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2941 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3001 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3061 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3121 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3181 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3241 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3301 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3361 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3721 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3781 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3841 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3901 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3961 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4021 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4081 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4141 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4201 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4261 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4321 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4381 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4441 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4501 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4561 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4621 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4681 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4741 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4801 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4861 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4921 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4981 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5041 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5281 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5761 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5821 ttctgtggat aaccgtatta ccgccatgca t
//
