LOCUS       sequence-290141-        5134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPAN3_245-550.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5134)
  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 5134)
  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_245-550_U
FEATURES             Location/Qualifiers
     source          1..5134
                     /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    2468..2589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2505..2524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2559..2578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2596..3051)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2733..2754)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2945..2964
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3078..3182
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3184..3541
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3184..3375
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3207..3227)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3392..3527
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3576..4370
                     /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             3576..4370
                     /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(3630..3649)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4240..4259
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4558..4577)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4602..4649
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4978..5134,1..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 ctatggctga tgaactccga caggagctga tcaacagaca
     1441 tttaataaca atggctcaaa ttgatcaagc agatatgcca gcagttccta cagaggttga
     1501 cagctaccat agcctattcc ctctagaacc actgccacct cccaaccgga tacagaaatc
     1561 aagtaatttt ggatatatta catcttgcta caaagctgta aacagcaaag atgatctgcc
     1621 atattgcctt cggaggatac atggttttcg tcttgttaac acaaagtgca tggtgttggt
     1681 cgacatgtgg aagaaaattc aacactcaaa tatcgtaact ttgcgtgaag tatttaccac
     1741 taaagcattt gctgagccct ctcttgtgtt tgcacatgat ttccatgctg gaggagaaac
     1801 tatgatgagc agacacttta atgaccctaa tgctgatgcc tacttcacca agagaaagtg
     1861 gggtcagcac gagggaccat tgcccaggca gcatgctgga ttattgccag aatctcttat
     1921 ttgggcatat attgtccaac taagttctgc attgcgtacc attcatacag caggtttggc
     1981 atgtcgagtt atggatccaa caaagattct gataactggc aaaacaaggt tgcgagtaaa
     2041 ttgtgttgga gtttttgatg ttttaacatt tgataacagt caaaataata atccattggc
     2101 attaatggcc cagtaccagc aagcagatct gatatcatta ggaaaagttg tgttggcttt
     2161 ggcttgcaac tctttggcag gaattcagcg agagaattta cagaaagcca tggaactggt
     2221 gacaatcaac tattcctctg acctgaagaa tctgattttg tatttgttga ctgaccaaaa
     2281 caggatgcga agtgtaaatg acatcatgcc catgattggt tagggtaccg cgggcccggg
     2341 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2401 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2461 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2581 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2641 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2701 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2761 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2821 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2881 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2941 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3001 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3061 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3121 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3181 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3361 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3421 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3481 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3541 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3601 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3661 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3721 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3781 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3841 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3901 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3961 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4021 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4081 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4141 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4201 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4261 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4321 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4381 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4441 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4501 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4561 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4621 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4681 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4741 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4801 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4861 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4921 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4981 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5041 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5101 cagagcgcag ataccaaata ctgtccttct agtg
//
