LOCUS       sequence-401505-        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPABPC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  AUTHORS   Braun JE, Huntzinger E, Fauser M, Izaurralde E
  TITLE     GW182 proteins directly recruit cytoplasmic deadenylase complexes to
            miRNA targets.
  JOURNAL   Mol Cell. 2011 Oct 7;44(1):120-33.
  PUBMED    21981923
REFERENCE   2  (bases 1 to 6656)
  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: "Mol Cell.";
            date: "2011-10-7"; volume: "44(1)"; pages: "120-33"
COMMENT     Sequence Label: pN3-HsPABPC1-V5_L
FEATURES             Location/Qualifiers
     source          1..6656
                     /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"
     CDS             2542..2583
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2596..2605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2602..2604,2605..2607,2608..3321)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2602..2604,2605..2607,2608..3318)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2647..2668)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2908..2927)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3255..3276
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3444..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3535..3554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3572..4027)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3709..3730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3921..3940
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4054..4158
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4160..4517
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4160..4351
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4183..4203)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4368..4503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4552..5346
                     /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             4552..5346
                     /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(4606..4625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5216..5235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5534..5553)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5578..5625
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5954..6542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6443..6462
                     /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 gcagagctgg tttagtgaac cgtcagatcc gctagcaatt
      601 gcggccacca tgaaccccag tgcccccagc taccccatgg cctcgctcta cgtgggggac
      661 ctccaccccg acgtgaccga ggcgatgctc tacgagaagt tcagcccggc cgggcccatc
      721 ctctccatcc gggtctgcag ggacatgatc acccgccgct ccttgggcta cgcgtatgtg
      781 aacttccagc agccggcgga cgcggagcgt gctttggaca ccatgaattt tgatgttata
      841 aagggcaagc cagtacgcat catgtggtct cagcgtgatc catcacttcg caaaagtgga
      901 gtaggcaaca tattcattaa aaatctggac aaatccattg ataataaagc actgtatgat
      961 acattttctg cttttggtaa catcctttca tgtaaggtgg tttgtgatga aaatggttcc
     1021 aagggctatg gatttgtaca ctttgagacg caggaagcag ctgaaagagc tattgaaaaa
     1081 atgaatggaa tgctcctaaa tgatcgcaaa gtatttgttg gacgatttaa gtctcgtaaa
     1141 gaacgagaag ctgaacttgg agctagggca aaagaattca ccaatgttta catcaagaat
     1201 tttggagaag acatggatga tgagcgcctt aaggatctct ttggcaagtt tgggcctgcc
     1261 ttaagtgtga aagtaatgac tgatgaaagt ggaaaatcca aaggatttgg atttgtaagc
     1321 tttgaaaggc atgaagatgc acagaaagct gtggatgaga tgaacggaaa ggagctcaat
     1381 ggaaaacaaa tttatgttgg tcgagctcag aaaaaggtgg aacggcagac ggaacttaag
     1441 cgcaaatttg aacagatgaa acaagatagg atcaccagat accagggtgt taatctttat
     1501 gtgaaaaatc ttgatgatgg tattgatgat gaacgtctcc ggaaagagtt ttctccattt
     1561 ggtacaatca ctagtgcaaa ggttatgatg gagggtggtc gcagcaaagg gtttggtttt
     1621 gtatgtttct cctccccaga agaagccact aaagcagtta cagaaatgaa cggtagaatt
     1681 gtggccacaa agccattgta tgtagcttta gctcagcgca aagaagagcg ccaggctcac
     1741 ctcactaacc agtatatgca gagaatggca agtgtacgag ctgttcccaa ccctgtaatc
     1801 aacccctacc agccagcacc cccttcaggt tacttcatgg cagctatccc acagactcag
     1861 aaccgtgctg catactatcc tcctagccaa attgctcaac taagaccaag tcctcgctgg
     1921 actgctcagg gtgccagacc tcatccattc caaaatatgc ccggtgctat ccgcccagct
     1981 gctcctagac caccatttag tactatgaga ccagcttctt cacaggttcc acgagtcatg
     2041 tcaacacagc gtgttgctaa cacatcaaca cagacaatgg gtccacgtcc tgcagctgca
     2101 gccgctgcag ctactcctgc tgtccgcacc gttccacagt ataaatatgc tgcaggagtt
     2161 cgcaatcctc agcaacatct taatgcacag ccacaagtta caatgcaaca gcctgctgtt
     2221 catgtacaag gtcaggaacc tttgactgct tccatgttgg catctgcccc tcctcaagag
     2281 caaaagcaaa tgttgggtga acggctgttt cctcttattc aagccatgca ccctactctt
     2341 gctggtaaaa tcactggcat gttgttggag attgataatt cagaacttct tcatatgctc
     2401 gagtctccag agtcactccg ttctaaggtt gatgaagctg tagctgtact acaagcccac
     2461 caagctaaag aggctgccca gaaagcagtt aacagtgcca ccggtgttcc aactgttagc
     2521 ggccgcggtc acccattcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     2581 acgtagggat ccatcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2641 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2701 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2761 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2821 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2881 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2941 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3001 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3061 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3121 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3181 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3241 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3301 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     3361 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3421 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3481 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3541 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3601 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3661 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3721 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3781 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3841 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3901 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3961 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4021 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4141 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4441 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4501 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4561 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4621 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4681 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4741 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4801 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4861 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4921 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4981 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5041 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5101 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5161 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5221 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5281 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5341 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5401 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5461 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5521 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5581 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5641 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5701 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5761 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5821 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
