LOCUS       sequence-290143-        4627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPAN3_551-687.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4627)
  AUTHORS   Christie M, Boland A, Huntzinger E, Weichenrieder O, Izaurralde E
  TITLE     Structure of the PAN3 pseudokinase reveals the basis for 
            interactions with the PAN2 deadenylase and the GW182 proteins.
  JOURNAL   Mol Cell. 2013 Aug 8;51(3):360-73. doi: 
            10.1016/j.molcel.2013.07.011.
  PUBMED    23932717
REFERENCE   2  (bases 1 to 4627)
  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: 
            "10.1016/j.molcel.2013.07"; journalName: "Mol Cell"; date: 
            "2013-08-8- 8"; volume: "51"; issue: "3"; pages: "360-73"
COMMENT     Sequence Label: pT7-V5-SBP-C1-HsPAN3_551-687_U
FEATURES             Location/Qualifiers
     source          1..4627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     333..352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        607..910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        911..1114
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1065..1085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1159..1178
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1159..1177
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1205..1214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1214..1255
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1277..1390
                     /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    1961..2082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1998..2017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2052..2071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2089..2544)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2226..2247)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2438..2457
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2571..2675
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2677..3034
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2677..2868
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2700..2720)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2885..3020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3069..3863
                     /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             3069..3863
                     /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(3123..3142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3733..3752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4051..4070)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4095..4142
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4471..4627,1..432)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
       61 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      541 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      601 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      661 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      721 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      781 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      841 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      901 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      961 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     1021 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     1081 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
     1141 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctc tagcgctacc
     1201 ggtcgccacc atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcccgg
     1261 aagcggtagc ggtacgatgg acgagaagac caccggctgg gggggcggcc acgtggtgga
     1321 gggcctggcc ggcgagctgg agcagctgcg ggccaggctg gagcaccacc ctcagggcca
     1381 gcgggagccc agatctcgag ctgctcgatt ttatactcaa ttggatgctg ctcaaatgag
     1441 aaatgatgtc atagaggaag accttgcaaa ggaggttcaa aatggaagac tgtttaggct
     1501 cctagcaaaa ttgggaacaa tcaatgagag gccggagttt cagaaggatc ccacttggtc
     1561 agagactgga gaccgttatc tgttgaaact ctttagggat catctttttc atcaggtgac
     1621 agaagcaggt gctccctgga ttgacctcag tcatataatt tcttgtctta acaagctaga
     1681 tgctggtgtg ccagaaaaaa taagcctgat ttccagagat gagaagagtg tacttgtggt
     1741 gacctacagt gacttaaagc gctgctttga aaatactttt caagaactga ttgcagctgc
     1801 aaatggtcag ttgtagggta ccgcgggccc gggatccacc ggatctagat aactgatcat
     1861 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1921 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1981 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2041 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2101 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2161 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2221 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2281 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2341 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2401 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2461 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2521 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2581 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2641 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2701 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2761 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2881 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2941 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3001 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3061 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3121 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3181 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3241 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3301 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3361 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3421 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3481 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3541 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3601 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3661 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3721 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3781 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3841 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3901 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3961 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4021 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4081 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4141 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4201 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4261 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4321 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4381 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4441 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4501 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4561 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4621 tctagtg
//
