LOCUS       sequence-290142-        5545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPAN3_245-687.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5545)
  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 5545)
  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_245-687_U
FEATURES             Location/Qualifiers
     source          1..5545
                     /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             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /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    2333..2454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2370..2389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2424..2443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2461..2916)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2598..2619)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2810..2829
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2943..3047
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3049..3406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3049..3240
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3072..3092)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3257..3392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3441..4235
                     /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             3441..4235
                     /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(3495..3514)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4105..4124
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4423..4442)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4467..4514
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4843..5431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5332..5351
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctatgg ctgatgaact ccgacaggag ctgatcaaca gacatttaat
      901 aacaatggct caaattgatc aagcagatat gccagcagtt cctacagagg ttgacagcta
      961 ccatagccta ttccctctag aaccactgcc acctcccaac cggatacaga aatcaagtaa
     1021 ttttggatat attacatctt gctacaaagc tgtaaacagc aaagatgatc tgccatattg
     1081 ccttcggagg atacatggtt ttcgtcttgt taacacaaag tgcatggtgt tggtcgacat
     1141 gtggaagaaa attcaacact caaatatcgt aactttgcgt gaagtattta ccactaaagc
     1201 atttgctgag ccctctcttg tgtttgcaca tgatttccat gctggaggag aaactatgat
     1261 gagcagacac tttaatgacc ctaatgctga tgcctacttc accaagagaa agtggggtca
     1321 gcacgaggga ccattgccca ggcagcatgc tggattattg ccagaatctc ttatttgggc
     1381 atatattgtc caactaagtt ctgcattgcg taccattcat acagcaggtt tggcatgtcg
     1441 agttatggat ccaacaaaga ttctgataac tggcaaaaca aggttgcgag taaattgtgt
     1501 tggagttttt gatgttttaa catttgataa cagtcaaaat aataatccat tggcattaat
     1561 ggcccagtac cagcaagcag atctgatatc attaggaaaa gttgtgttgg ctttggcttg
     1621 caactctttg gcaggaattc agcgagagaa tttacagaaa gccatggaac tggtgacaat
     1681 caactattcc tctgacctga agaatctgat tttgtatttg ttgactgacc aaaacaggat
     1741 gcgaagtgta aatgacatca tgcccatgat tggtgctcga ttttatactc aattggatgc
     1801 tgctcaaatg agaaatgatg tcatagagga agaccttgca aaggaggttc aaaatggaag
     1861 actgtttagg ctcctagcaa aattgggaac aatcaatgag aggccggagt ttcagaagga
     1921 tcccacttgg tcagagactg gagaccgtta tctgttgaaa ctctttaggg atcatctttt
     1981 tcatcaggtg acagaagcag gtgctccctg gattgacctc agtcatataa tttcttgtct
     2041 taacaagcta gatgctggtg tgccagaaaa aataagcctg atttccagag atgagaagag
     2101 tgtacttgtg gtgacctaca gtgacttaaa gcgctgcttt gaaaatactt ttcaagaact
     2161 gattgcagct gcaaatggtc agttgtaggg taccgcgggc ccgggatcca ccggatctag
     2221 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2281 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2461 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2521 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2581 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2641 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2701 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2761 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2821 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2881 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     2941 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3001 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3061 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3241 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3361 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3421 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3481 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3541 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3601 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3661 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3721 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3781 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3841 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3901 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3961 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4021 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4081 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4141 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4201 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4261 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4321 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4381 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4441 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4501 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4561 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4621 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4681 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4741 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4981 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5461 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5521 ggataaccgt attaccgcca tgcat
//
