LOCUS       sequence-422631-        6277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPAN3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  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 6277)
  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: pT7-V5-SBP-C1-HsPAN3_M
FEATURES             Location/Qualifiers
     source          1..6277
                     /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             608..649
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             671..784
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    3005..3126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3042..3061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3096..3115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3133..3588)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3270..3291)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3482..3501
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3615..3719
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3721..4078
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3721..3912
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3744..3764)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3929..4064
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4113..4907
                     /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             4113..4907
                     /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(4167..4186)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4777..4796
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5095..5114)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5139..5186
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5515..6103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6004..6023
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      661 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      721 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      781 gcccagatct cgagctatgg atggaggtgc tttaactgat acaagtctca cagattccta
      841 ttttagcacc agctttattg gagtcaatgg atttggaagc cctgtagaaa caaaatatcc
      901 cctgatgcag agaatgacta atagtagcag ctccccaagc cttctaaatg acagtgccaa
      961 gccatattca gcccatgatc ctctaacatc acctgcttca tccttgttta atgactttgg
     1021 tgccctcaac atctctcaga gacgaaagac accaaatcct actgcaagcg agtttattcc
     1081 taaaggagga tcaacctcca ggctgagtaa cgtgtcccag tcaaatatgt ctgccttctc
     1141 tcaagttttc tctcacccat ccatgggaag ccctgctact gctggattag cgccaggaat
     1201 gtcgttgtct gctgggtctt cccctcttca ttcccccaag attactccac atacttctcc
     1261 tgctcccaga agaagaagtc acactccaaa tccagcaagt tacatggtgc cttctagtgc
     1321 ctctacatct gttaataatc ctgtttctca gactccgtct tctggtcagg tgatccaaaa
     1381 ggaaactgtt ggtgggacga cttacttcta tacagacaca actccagcac ctttgactgg
     1441 aatggtgttt ccaaactatc atatttatcc tccaactgca cctcacgttg cttatatgca
     1501 accgaaagca aacgcacctt ccttcttcat ggctgatgaa ctccgacagg agctgatcaa
     1561 cagacattta ataacaatgg ctcaaattga tcaagcagat atgccagcag ttcctacaga
     1621 ggttgacagc taccatagcc tattccctct agaaccactg ccacctccca accggataca
     1681 gaaatcaagt aattttggat atattacatc ttgctacaaa gctgtaaaca gcaaagatga
     1741 tctgccatat tgccttcgga ggatacatgg ttttcgtctt gttaacacaa agtgcatggt
     1801 gttggtcgac atgtggaaga aaattcaaca ctcaaatatc gtaactttgc gtgaagtatt
     1861 taccactaaa gcatttgctg agccctctct tgtgtttgca catgatttcc atgctggagg
     1921 agaaactatg atgagcagac actttaatga ccctaatgct gatgcctact tcaccaagag
     1981 aaagtggggt cagcacgagg gaccattgcc caggcagcat gctggattat tgccagaatc
     2041 tcttatttgg gcatatattg tccaactaag ttctgcattg cgtaccattc atacagcagg
     2101 tttggcatgt cgagttatgg atccaacaaa gattctgata actggcaaaa caaggttgcg
     2161 agtaaattgt gttggagttt ttgatgtttt aacatttgat aacagtcaaa ataataatcc
     2221 attggcatta atggcccagt accagcaagc agatctgata tcattaggaa aagttgtgtt
     2281 ggctttggct tgcaactctt tggcaggaat tcagcgagag aatttacaga aagccatgga
     2341 actggtgaca atcaactatt cctctgacct gaagaatctg attttgtatt tgttgactga
     2401 ccaaaacagg atgcgaagtg taaatgacat catgcccatg attggtgctc gattttatac
     2461 tcaattggat gctgctcaaa tgagaaatga tgtcatagag gaagaccttg caaaggaggt
     2521 tcaaaatgga agactgttta ggctcctagc aaaattggga acaatcaatg agaggccgga
     2581 gtttcagaag gatcccactt ggtcagagac tggagaccgt tatctgttga aactctttag
     2641 ggatcatctt tttcatcagg tgacagaagc aggtgctccc tggattgacc tcagtcatat
     2701 aatttcttgt cttaacaagc tagatgctgg tgtgccagaa aaaataagcc tgatttccag
     2761 agatgagaag agtgtacttg tggtgaccta cagtgactta aagcgctgct ttgaaaatac
     2821 ttttcaagaa ctgattgcag ctgcaaatgg tcagttgtag ggtaccgcgg gcccgggatc
     2881 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2941 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3001 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3061 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3121 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3181 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3241 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3301 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3361 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3421 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3481 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3541 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3721 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3781 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3841 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3901 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3961 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4021 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4081 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4141 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4201 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4261 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4321 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4381 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4441 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4501 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4561 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4621 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4681 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4741 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4801 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4861 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4921 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5041 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5101 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5161 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5221 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5281 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5341 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5401 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5461 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5521 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5581 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5641 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5701 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5761 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5821 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5881 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5941 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6001 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6061 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6121 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6181 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     6241 cggggtcatt agttcatagc ccatatatgg agttccg
//
