LOCUS       sequence-481981-        4707 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express Histone 3.3-SmBiT Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4707)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: CMV/H3.3 SmBiT Vector
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    147..430
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        621..1000
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1001..1204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1155..1175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1340..1472
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1455..1474
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1516..1535
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1516..1534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2133..2151)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2188..2309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2199..2218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2253..2272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2554..2911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2554..2745
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2577..2597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2762..2897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2926..2973
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3005..3799
                     /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(3059..3078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3669..3688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3863..3911
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4230..4249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
       61 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      121 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
      181 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
      241 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
      301 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
      361 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
      421 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
      481 tcttcaatat tggccattag ccatattatt cattggttat atagcataaa tcaatattgg
      541 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
      601 tccaatatga ccgccatgtt ggcattgatt attgactagt tattaatagt aatcaattac
      661 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      721 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      781 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      841 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
      901 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
      961 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1021 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1081 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     1141 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     1201 agctggttta gtgaaccgtc agatcactag aagctttatt gcggtagttt atcacagtta
     1261 aattgctaac gcagtcagtg cttctgacac aacagtctcg aacttaagct gcagaagttg
     1321 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     1381 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     1441 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
     1501 ttaaggctag agtattaata cgactcacta tagggctagc gatcgccatg gctcgtacaa
     1561 agcagactgc ccgcaaatcg accggtggta aagcacccag gaagcaactg gctacaaaag
     1621 ccgctcgcaa gagtgcgccc tctactggag gggtgaagaa acctcatcgt tacaggcctg
     1681 gtactgtggc gctccgtgaa attagacgtt atcagaagtc cactgaactt ctgattcgca
     1741 aacttccctt ccagcgtctg gtgcgagaaa ttgctcagga ctttaaaaca gatctgcgct
     1801 tccagagcgc agctatcggt gctttgcagg aggcaagtga ggcctatctg gttggccttt
     1861 ttgaagacac caacctgtgt gctatccatg ccaaacgtgt aacaattatg ccaaaagaca
     1921 tccagctagc acgccgcata cgtggagaac gtgctgtttc tcaaggcagt tcaggtggtg
     1981 gcgggagcgg aggtggaggc tcgagcggtg tgaccggcta ccggctgttc gaggagattc
     2041 tgtaaggccg cgactctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     2101 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2161 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2221 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2281 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2341 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     2401 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     2461 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     2521 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     2581 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2641 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2701 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2761 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2821 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2881 aggctttttt ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca
     2941 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     3001 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3061 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3181 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3301 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3781 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3841 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     3901 ttttttgtgt gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca
     3961 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     4021 cccggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4561 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4681 tagcggtggt ttttttgttt gcaagca
//
