LOCUS       sequence-94268-e        8671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express H3.3 chaperone HIRA in mammalian cells (CMV promoter). Also 
            suitable for in vitro transcription (T7 promoter) for injecting mRNA
            into oocytes. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8671)
  AUTHORS   Inoue A, Zhang Y
  TITLE     Nucleosome assembly is required for nuclear pore complex assembly in
            mouse zygotes.
  JOURNAL   Nat Struct Mol Biol. 2014 Jul;21(7):609-16. doi: 10.1038/nsmb.2839. 
            Epub 2014 Jun 8.
  PUBMED    24908396
REFERENCE   2  (bases 1 to 8671)
  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: "10.1038/nsmb.2839";
            journalName: "Nat Struct Mol Biol"; date: "2014-07"; volume: "21"; 
            issue: "7"; pages: "609-16"
COMMENT     Sequence Label: pcDNA-HIRA-polyA
FEATURES             Location/Qualifiers
     source          1..8671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4174..4203
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4219..4236
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4259..4276)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4265..4489
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4535..4963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4622..4641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4832..4853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4972..4992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4977..5307
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5158..5293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5220..5239
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5374..6168
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5428..5447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6038..6057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6344..6465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6381..6400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6435..6454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6514..6530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6514..6530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6527..6549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6538..6554
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6562..6568,6569..6586,6587..6592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6607..6628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6745..6762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6916..7504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6996..7015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7675..8466,8467..8535))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8298..8317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8536..8640)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttcga attcatgaag ctcttgaagc caacctgggt caaccacaat gggaagccaa
      961 ttttttcagt tgatattcac cctgatggga ccaagtttgc aactggagga caagggcagg
     1021 attctgggaa ggttgtgatc tggaatatgt ctccagtcct ccaggaggat gacgagaagg
     1081 atgaaaatat tcccaagatg ctttgccaga tggacaatca cttagcatgt gtgaactgtg
     1141 tgcggtggtc aaacagtggg atgtatttag cttctggggg agatgacaaa ctgattatgg
     1201 tgtggaagcg ggctacgtac attgggccca gcactgtgtt tggttccagt ggtaagcttg
     1261 ccaatgtgga gcaatggcgg tgtgtctcca tcctccggag tcactcaggc gatgtgatgg
     1321 atgtagcatg gtctccccac gacgcctggc tggcctcatg cagcgtggat aacactgttg
     1381 tcatttggaa tgccgtgaag ttcccagaaa ttcttgcaac tctgagaggt cattctggct
     1441 tagtaaaagg tttgacttgg gatcccgttg gtaaatatat tgcctctcaa gctgatgatc
     1501 gaagtttgaa ggtatggagg acgctggact ggcagctaga gactagcatc accaagcctt
     1561 ttgatgagtg tggaggaacg acgcatgttc tccggcttag ttggtcacct gatggccatt
     1621 acctggtatc tgcccatgcc atgaataatt ctggccccac tgctcagatc atcgaaagag
     1681 agggctggaa gaccaacatg gactttgtgg gtcaccggaa agctgtgact gttgtgaaat
     1741 tcaacccaaa aatcttcaag aagaagcaga agaatgggag ctctacaaag cccagctgcc
     1801 catactgctg ctgtgctgtt ggcagcaagg accgctcact ctctgtctgg ctcacatgtt
     1861 tgaaacggcc tctggttgtc atccatgaac tgtttgacaa gtccatcatg gatatttcct
     1921 ggactctgaa tgggttgggt atcctggtgt gctccatgga cggctctgtg gcgttccttg
     1981 atttctctca ggatgaactc ggagaccccc tgagtgagga ggaaaagagc cgaattcacc
     2041 agtctaccta tggcaagagc ctggcaataa tgactgaggc ccagctttcc acagctgtta
     2101 ttgagaaccc tgagatgctc aagtaccagc ggagacagca acagcagcag ctggatcaga
     2161 agaatgccac tactagggag acaagctcag catcctcagt cacgggtgtg gtcaatgggg
     2221 aaagtctaga agatatcaga aagaatcttt tgaagaaaca agtggaaact cggacagcag
     2281 atggtcggag gagaatcacg cctctttgca tagcacagct ggacactggg gacttctcca
     2341 cggcattctt caacagcatc ccactctcca gctccctagc aggcaccatg ctctcctctc
     2401 ctagtggtca gcagctacta ccactggact ccagtacccc ctccttcggc gcctcaaagc
     2461 cttgcacaga accagtggca gccaccagtg ccaggcctac aggcgaatct gtcagtaagg
     2521 acagtatgaa tgctacctct actcctgctg catcgtcacc ctctgtgtta acaaccccat
     2581 ccaagattga acccatgaaa gcatttgatt cccggttcac agaacggtcc aaagccacac
     2641 caggtgctcc ttccttgacc agtgtgattc caacagctgt tgaaaggttg aaagagcaga
     2701 acctcgtcaa ggagctgagg tcccgggaac tggagagcag cagtgacagc gatgagaagg
     2761 tccacctagc caagccctct tcactgtcca agcgcaaact tgagcttgag gtagagacgg
     2821 tggaaaagaa gaagaaaggt cgccctagga aggattcacg tcttttaccc atgtctctgt
     2881 ccgtccagtc tccagctgcc ctgtctacag agaaggaggc catgtgtctg tctgcaccag
     2941 cacttgcact gaagctgcca attccaggcc cacagagagc gttcaccctc caggtgagct
     3001 ctgacccctc catgtacatt gaggtggaga atgaagtgac caccgttggg gggataaggc
     3061 tgagtcgcct gaagtgcaac cgtgaaggga aggagtggga gacagtgctc agcagtcgcg
     3121 tcctcactgc tgccggcagc tgtgatgtgg tatgtgttgc ctgtgaaaag aggatgctgt
     3181 cggtgttctc tacctgtggt cgccgtctcc tccctcccat cctccttcca tctccaatct
     3241 ctactttgca ctgcacgggc ccctacgtca tggcactcac cgctgcagcc acactgtctg
     3301 tctgggatgt tcacagacag gtggttgtgg tgaaagaaga atctctacac tccatcttgt
     3361 caggaagtga tatgacggtg tcacagatct tgctaacaca gcatggaatc ccagtgatga
     3421 atttgtctga tgggaaagca tactgcttta atccatccct ctccacatgg aacctggttt
     3481 ctgacaagca agattcattg gcccagtgtg cagacttcag gaacagcctg ccgtcccagg
     3541 atgccatgct ttgttcagga ccgttagcca taattcaggg ccgcacctcc aactctggaa
     3601 ggcaagctgc ccggctcttc tccgtgcctc atgtggtaca gcaggagacc acactggcct
     3661 acctagagaa ccaggttgct gcggcactta ccctgcagtc aagccacgag tatcgtcact
     3721 ggctcctcct ttatgctcgg taccttgtga atgaagggtt tgaataccgc ctccgtgaaa
     3781 tatgcaagga cttgctgggt ccagttcact gctccactgg aagtcagtgg gagtcaacag
     3841 tagtgggtct gcggaaaagg gagctgctga aggaactgtt gccagtcatt gggcagaatc
     3901 ttcgattcca gcgcctcttt actgagtgcc aggaacaact ggacatcctg agagacaagt
     3961 aggcggccgc tcgactatgg gcaccaaaga acctgtaaac gttatctttt taaattgaat
     4021 gtgcacaaat aaaagtttgg aaaagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4081 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaata cgcatcatac
     4141 gctgtggcgt acctcgagtc tagagggccc ttcgaacaaa aactcatctc agaagaggat
     4201 ctgaatatgc ataccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     4261 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4321 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4381 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4441 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4501 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4681 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4741 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4801 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4861 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4921 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4981 gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg
     5041 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5101 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5161 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5221 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5281 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5341 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5401 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5461 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5521 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5581 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5641 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5701 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5761 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5821 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5881 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5941 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6001 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6061 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6121 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6181 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6241 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6301 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6481 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6541 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6601 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6661 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6721 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6781 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6841 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6901 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6961 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7021 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7081 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     7141 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7201 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7261 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7321 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7381 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7441 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7501 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7561 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7621 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7681 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7741 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7801 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7861 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7921 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7981 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8041 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8101 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8161 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8221 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8281 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8341 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8401 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8461 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8521 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8581 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8641 cacatttccc cgaaaagtgc cacctgacgt c
//
