LOCUS       sequence-401023-        6373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsXRN1_1174-1706.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   Braun JE, Truffault V, Boland A, Huntzinger E, Chang CT, Haas G, 
            Weichenrieder O, Coles M, Izaurralde E
  TITLE     A direct interaction between DCP1 and XRN1 couples mRNA decapping to
            5' exonucleolytic degradation.
  JOURNAL   Nat Struct Mol Biol. 2012 Dec;19(12):1324-31. doi: 
            10.1038/nsmb.2413. Epub 2012 Nov 11.
  PUBMED    23142987
REFERENCE   2  (bases 1 to 6373)
  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"; doi: "10.1038/nsmb.2413";
            journalName: "Nat Struct Mol Biol"; date: "2012-12"; volume: "19"; 
            issue: "12"; pages: "1324-31"
COMMENT     Sequence Label: pT7-EGFP-C1-HsXRN1_1174-1706_K
FEATURES             Location/Qualifiers
     source          1..6373
                     /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    3161..3282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3252..3271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3289..3744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3426..3447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3638..3657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3771..3875
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3877..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3877..4068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4269..5063
                     /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             4269..5063
                     /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(4323..4342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4933..4952
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5251..5270)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5295..5342
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5671..6259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6160..6179
                     /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 ctcaagcttc gaattctgca gtcgacatgc atgggagtcg
     1441 ctctgaaact ggaaatcaga agttgacagc catcgtaaaa ccacaaccag ctgtacatca
     1501 acatagctca agttcatcag tttcctctgg gcatttggga gccctcaacc attcccctca
     1561 atcacttttt gttcctactc aagtacctac taaagatgat gatgaattct gcaacatttg
     1621 gcagtcctta cagggatctg gaaagatgca atactttcag ccaactatac aagagaaggg
     1681 tgcagttcta cctcaagaaa taagccaagt aaatcaacat cataaatctg gctttaatga
     1741 caacagtgtt aaatatcagc aaagaaaaca tgaccctcac agaaaattta aagaagagtg
     1801 taagagtcct aaagctgagt gttggtccca aaaaatgtcc aataagcagc ctaactctgg
     1861 aattgagaac tttttagcat ctttgaatat ctccaaagaa aatgaagtac agtcatctca
     1921 tcatggggag cctccaagtg aagagcattt gtcaccacag tcatttgcca tgggaacacg
     1981 gatgcttaaa gaaattctaa aaattgatgg ctctaacact gtggaccata agaatgaaat
     2041 caaacagatt gctaatgaaa tccctgtttc ctctaacaga agagatgaat atggattacc
     2101 ctctcagcct aaacaaaata agaaattagc atcttatatg aacaagcctc acagtgctaa
     2161 tgagtaccat aatgttcagt ctatggacaa tatgtgttgg cctgccccca gccagatccc
     2221 tcctgtatcc acaccagtaa ctgaactttc tcgaatttgt tcccttgttg gaatgccaca
     2281 acctgatttc tcctttctta ggatgccaca gacaatgacc gtttgccaag taaaattatc
     2341 taatggctta ctggtacatg ggccacagtg ccactctgaa aatgaagcca aagagaaagc
     2401 tgcacttttt gctttacaac agttgggctc cttaggcatg aatttccctt tgccttcaca
     2461 agtatttgca aattatcctt cagctgtacc acctggaacc attcctccag cctttccccc
     2521 acctactggc tgggatcact atggaagcaa ctatgcattg ggggcagcta atataatgcc
     2581 ttcgtcgtct catctctttg gctcaatgcc atggggacca tcggtgccag ttcctgggaa
     2641 gcccttccat catactttat attctgggac catgcccatg gctgggggaa taccaggggg
     2701 tgtgcacaat cagtttatac ctctgcaggt tactaaaaaa agggttgcaa acaaaaagaa
     2761 ctttgagaat aaggaagccc agagttctca agccactcca gttcagacta gccagccaga
     2821 ttcttccaac attgtcaaag taagtccacg ggagagctca tcagcttctt tgaagtcctc
     2881 tccgattgct caacctgcat cttcttttca agttgaaact gcctctcaag gccatagtat
     2941 atctcaccat aagtcaacac caatctcttc ttcaagaaga aaatcaagaa aactggctgt
     3001 taattttggt gtttctaaac cttctgagta aggatccacc ggatctagat aactgatcat
     3061 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3121 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3181 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3241 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3301 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3361 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3421 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3481 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3541 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3601 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3661 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3721 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3841 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4261 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4321 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4381 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4441 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4501 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4561 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4621 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4681 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4741 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4801 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4861 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4921 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4981 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5041 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5101 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5221 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5281 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5341 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5401 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5461 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6361 taccgccatg cat
//
