LOCUS       sequence-289132-        6691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1-del399-516.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  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 6691)
  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-del399-516_J
FEATURES             Location/Qualifiers
     source          1..6691
                     /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    3479..3600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3516..3535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3570..3589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3607..4062)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3744..3765)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3956..3975
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4089..4193
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4195..4552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4195..4386
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4218..4238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4403..4538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4587..5381
                     /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             4587..5381
                     /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(4641..4660)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5251..5270
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5569..5588)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5613..5660
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5989..6577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6478..6497
                     /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 cgaaaagaca agaggagaaa aacccttgtt ataattgaga aaacctacag
     2641 cttactcctt gatgtggagg actatgaaag acgttatctc ctaagtctgg aagaagagcg
     2701 acctgcccta atggatgaca gaaagcacaa aatttgtagc atgtatgaca acttaagggg
     2761 gaaattgcct ggacaagaga ggcctagtga tgaccacttt gtacagatca tgtgtatccg
     2821 aaaagggaag agaatggttg cccgtattct tcctttcctc tccacagagc aagcagctga
     2881 cattctcatg acaacagcca ggaacctccc tttccttatc aagaaggatg cacaagatga
     2941 ggtgctgcca tgcttactga gtcccttctc tctccttctc tatcatcttc catcagtgag
     3001 tatcaccagc cttttgcgac agctaatgaa cctacctcaa agtgcagcta caccagcact
     3061 ctccaatcct cacctcactg ctgtgctcca gaacaagttt ggcctgtcac tgctcctcat
     3121 cctcctgagc cgtggtgaag acctacagag ttcagaccct gctacagaat caacacaaaa
     3181 taatcagtgg acggaggtga tgttcatggc aacacgagaa cttctgcgga ttccccaagc
     3241 agccctggcc aagccaatct ctatacctac aaacctagtg tccctctttt ctcgctatgt
     3301 tgaccggcag aaactgaact tgctggagac aaaactgcag ctagttcagg ggatacgata
     3361 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3421 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3601 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3661 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3721 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3781 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3841 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3901 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3961 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4021 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4201 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4321 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4381 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4441 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4501 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4561 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4621 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4681 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4741 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4801 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4861 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4921 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4981 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5041 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5101 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5161 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5221 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5281 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5341 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5401 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5461 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5521 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5581 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5641 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5701 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5761 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5821 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6121 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6601 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6661 ttctgtggat aaccgtatta ccgccatgca t
//
