LOCUS       sequence-393059-        8669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRRP12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8669)
  AUTHORS   Kashima I, Jonas S, Jayachandran U, Buchwald G, Conti E, Lupas AN, 
            Izaurralde E
  TITLE     SMG6 interacts with the exon junction complex via two conserved 
            EJC-binding motifs (EBMs) required for nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2010 Nov 1. 24(21):2440-50.
  PUBMED    20930030
REFERENCE   2  (bases 1 to 8669)
  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: "Genes Dev. 
            2010 Nov 1. 24(21):2440-50."
COMMENT     Sequence Label: pT7-EGFP-C1-HsRRP12_K
FEATURES             Location/Qualifiers
     source          1..8669
                     /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    5397..5518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5434..5453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5488..5507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5525..5980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5662..5683)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5874..5893
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6007..6111
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6113..6470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6113..6304
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6136..6156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6321..6456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6505..7299
                     /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             6505..7299
                     /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(6559..6578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7169..7188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7487..7506)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7531..7578
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7907..8495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8396..8415
                     /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 ctatgggtcg ctcgggaaag ttgccctctg gtgtctcagc
     1381 taagttgaag cgctggaaga aaggccacag cagcgacagc aaccccgcca tctgccgcca
     1441 ccgtcaggcc gcccgcagcc gcttcttcag ccggccgtca ggaaggagtg acctgacagt
     1501 cgatgctgtg aagttacata atgagctgca gtcagggtcc ttgcgcttgg gcaaaagcga
     1561 agccccggag acgcccatgg aagaagaggc ggagctggtt ctcaccgaga agtcctcggg
     1621 taccttcctg agtggccttt ccgactgcac aaacgtcacc ttcagcaaag tacagcgctt
     1681 ctgggagtcc aactcggctg cccacaagga gatctgtgct gttctggctg ctgtcactga
     1741 ggtgattcgc tcccagggag ggaaggagac ggagactgag tacttcgctg ctctgatgac
     1801 aacaatggaa gcagtggagt ccccggagtc cctggccgcc gttgcttacc tgctgaacct
     1861 tgtcctgaag cgtgttccca gccctgtgct tattaagaag ttctctgata cctccaaagc
     1921 cttcatggat atcatgtcag ctcaggccag cagcggctcc acctctgtcc tccgatgggt
     1981 cctttcctgc ctggccaccc ttctgcggaa gcaagacctg gaggcctggg gctaccccgt
     2041 gacccttcag gtgtaccatg ggctgctgag cttcacggtg catcccaagc ccaagatccg
     2101 gaaggctgcc cagcatggag tatgctcagt cctcaagggc agtgaattca tgtttgaaaa
     2161 ggcccctgcc catcatcctg ctgccatttc cactgccaag ttctgcatcc aggagattga
     2221 gaagtctgga ggctccaagg aggccaccac cacgctgcac atgctgacgc tgctgaagga
     2281 cctgctgccc tgcttcccgg aaggcctggt gaagagctgc agtgagactc tcctcagggt
     2341 catgaccttg agccatgtgc tggtgacagc ctgtgccatg caggcctttc acagcctctt
     2401 ccacgccagg cctggcctga gcaccctgtc agcagagctc aacgcccaga tcatcacggc
     2461 cctgtacgac tatgttccca gtgagaatga tttacaaccc ctgctagcct ggcttaaggt
     2521 catggagaaa gcccacatca acctggtgag gttgcagtgg gacctggggc taggccacct
     2581 ccctcgcttt tttggaactg cggtgacctg cctcctttcc ccacactcgc aagtgctgac
     2641 tgctgctacg cagagcctca aggagatcct gaaggaatgc gtggctcccc acatggctga
     2701 cattggctcc gtgacctcct cggcctcagg ccctgcccaa tctgttgcca agatgttcag
     2761 ggcagtggag gagggcctga cgtacaaatt ccatgcggcc tggagctccg tgttgcagct
     2821 gctgtgtgtc ttcttcgagg cgtgtgggag acaggcccac cctgtgatga ggaagtgcct
     2881 ccagtccctg tgtgacctgc gcctctcccc tcatttcccc cacacggcgg ctcttgacca
     2941 ggcagtgggg gctgcggtga ccagtatggg acctgaggtg gtgctgcagg ctgtgccttt
     3001 ggaaattgat ggctctgagg agactctgga tttcccacgg agctggctgc tgcctgtcat
     3061 ccgagaccat gttcaggaaa cgcgacttgg ttttttcacc acctacttct tgcccctggc
     3121 taacaccctg aagagcaaag ccatggacct ggctcaggca ggcagcacag tggaatctaa
     3181 gatctacgac acactccagt ggcagatgtg gacactcctg cctgggttct gcacaaggcc
     3241 tacagatgtg gccatctcct tcaaagggct ggcacggacg ctgggcatgg ccatcagcga
     3301 gcgtccagac ctgagggtca ccgtgtgcca ggccctgcgc accctcatca ccaagggctg
     3361 ccaggcagag gctgaccgtg ctgaagtgag tcgctttgcc aagaactttc tgccgatcct
     3421 cttcaacctg tatgggcagc ccgtggcagc cggggacact ccagcccctc gccgggctgt
     3481 gctggaaacc atcagaactt acctcaccat cactgacact cagttggtga acagtctcct
     3541 ggaaaaagcc agtgagaagg tgctcgaccc tgccagctct gactttacca gattgtctgt
     3601 cctggacctg gtcgtggcct tggctccgtg tgctgacgaa gctgccatca gtaagctata
     3661 ctccaccatc cggccctacc tagagagcaa ggcccacggg gtgcagaaga aggcctaccg
     3721 agtgctggag gaggtgtgtg ccagtcctca gggccccggg gccctcttcg tgcagagcca
     3781 cctggaggac ctgaagaaga cactgctgga ctcgctgcgg agcacctcct cacccgccaa
     3841 gaggccccgt ttgaagtgcc tcctacacat cgtgaggaag ctctcagctg aacacaagga
     3901 gttcatcact gccctcatcc cagaggtgat cctgtgcacc aaggaggtgt cggtgggcgc
     3961 acggaagaac gcttttgcac tgctcgtgga gatgggccat gctttcctaa ggtttggctc
     4021 gaaccaggaa gaggccctgc agtgctacct cgtcctgatc taccctggcc tggtgggcgc
     4081 ggtgaccatg gtcagctgca gcatcctggc cctgacccac ctccttttcg agtttaaagg
     4141 tctgatgggg accagtacag tggagcagct gctggagaat gtgtgcctgc ttctggcctc
     4201 ccgcacccgt gacgtggtca agtctgcact gggcttcatc aaggtggcag tgactgtcat
     4261 ggacgtggcg cacctggcca aacatgtgca gctggtgatg gaagccattg ggaagctttc
     4321 agatgacatg cggcggcact tccgcatgaa gcttcggaac ctgttcacca agttcatccg
     4381 caagtttgga tttgagctgg tgaaaaggct gttgcccgag gagtaccaca gagtcctggt
     4441 caacatccgg aaagctgagg cccgggccaa gaggcaccga gccctgagcc aggctgccgt
     4501 ggaggaggaa gaagaggagg aggaggagga ggagcccgcc cagggcaaag gtgacagcat
     4561 tgaggagatt ttagctgact cagaggacga ggaggacaat gaggaggagg aaagaagccg
     4621 aggcaaggag cagcggaagc tggcacgaca gaggagccgg gcatggctga aagagggcgg
     4681 tggggacgag cccctcaact tcctggatcc caaggtggcc caacgagtcc tggccacgca
     4741 gccagggcca ggccggggca ggaagaagga ccacagcttc aaggtgagcg ccgatggccg
     4801 gctgatcata agggaggagg cagacggcaa caagatggag gaagaggaag gtgccaaagg
     4861 cgaagatgaa gagatggctg acccaatgga agatgtgatc atcaggaata aaaagcacca
     4921 gaagctcaag caccagaaag aggctgagga ggaggagctg gagatacccc ctcagtacca
     4981 agctggaggc tctggcattc atcgccctgt ggccaagaag gctatgcctg gggctgaata
     5041 caaggccaag aaagcaaaag gtgatgtgaa gaagaaaggc cggccggatc cctatgccta
     5101 catccccctc aacagaagca agctcaaccg caggaagaag atgaagctgc agggacagtt
     5161 caaaggcctg gtgaaggctg cccggcgagg ttcccaggtg ggacacaaaa accgcagaaa
     5221 ggatcgtcga ccctgacaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     5281 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     5341 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     5401 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5461 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     5521 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5581 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5641 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5701 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5761 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5821 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5881 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5941 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     6001 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6061 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     6121 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     6181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     6481 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6541 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6601 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6661 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     6721 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6781 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6841 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6901 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6961 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     7021 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     7081 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     7141 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     7201 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     7261 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     7321 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7381 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7441 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     7501 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     7561 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     7621 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     7681 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     7741 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7801 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7861 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7921 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7981 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8041 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8101 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8161 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8221 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8281 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8341 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8401 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8461 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8521 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8581 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     8641 ttagttcata gcccatatat ggagttccg
//
