LOCUS       sequence-418360-        5734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid expressing H3.3 Wt histone in tandem with BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5734)
  TITLE     Castro lab plasmids Sep 2023
REFERENCE   2  (bases 1 to 5734)
  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"
COMMENT     Sequence Label: pIRES-H3.3WT-BFP
FEATURES             Location/Qualifiers
     source          1..5734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             159..953
                     /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             159..953
                     /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(213..232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     823..842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1141..1160)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1185..1232
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1561..2149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2050..2069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2324..2627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2628..2831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2782..2802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3392..3941
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3536..3553)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3763..3782
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3944..3946,3947..3949,3950..4663)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(3989..4010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    4785..4906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4822..4841)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4876..4895
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4913..5368)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5050..5071)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5262..5281
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5395..5499
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5501..5734,1..124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5501..5692
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5524..5544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5709..5734,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
       61 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      121 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      181 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      241 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      301 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
      361 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      421 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      481 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      541 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      601 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      661 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
      721 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
      781 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
      841 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
      901 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
      961 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1021 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1081 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     1141 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     1201 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     1261 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     1321 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     1381 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     1441 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1501 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1561 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1621 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1681 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1741 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1801 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1861 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1921 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1981 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2041 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2101 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2161 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2221 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     2281 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2341 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2401 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2461 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2521 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2581 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     2641 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2701 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2761 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2821 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
     2881 gctcaccatg actagtgccc gaaccaagca gactgctcgt aagtccaccg gtgggaaagc
     2941 cccccgcaaa cagctggcca cgaaagccgc caggaaaagc gctccctcta ccggcggggt
     3001 gaagaagcct catcgctaca ggcccgggac cgtggcgctt cgagagattc gtcgttatca
     3061 gaagtcgacc gagctgctca tccggaagct gcccttccag aggttggtga gggagatcgc
     3121 gcaggatttc aaaaccgacc tgaggtttca gagcgcagcc atcggtgcgc tgcaggaggc
     3181 tagcgaagcg tacctggtgg gtctgttcga agataccaac ctgtgtgcca tccacgctaa
     3241 gagagtcacc atcatgccca aagacatcca gttggctcgc cggatacggg gagagagagc
     3301 ttaagaagga tccctgcagt tcctgcagtc gacggtaccg cgggcccggg atcccaattg
     3361 ctcgagcggg atcaattccg ccccccccct aacgttactg gccgaagccg cttggaataa
     3421 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3481 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3541 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3601 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     3661 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     3721 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3781 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3841 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     3901 aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggtga gcaagggcga
     3961 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4021 caagttcagc gtgaggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     4081 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgag
     4141 ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4201 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcac
     4261 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4321 gaagggcgtc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     4381 caacagccac aacatctata tcatggccgt caagcagaag aacggcatca aggtgaactt
     4441 caagatccgc cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4501 cacccccatc ggcgacggcc ccgtgctgct gcccgacagc cactacctga gcacccagtc
     4561 cgtgctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttccgcac
     4621 cgccgccggg atcactctcg gcatggacga gctgtacaag taagcggccg cgactctaga
     4681 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4741 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4801 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4861 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4921 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4981 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5041 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5101 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5161 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5221 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5281 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5341 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5401 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5461 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5701 ctccgcccat cccgccccta actccgccca gttc
//
