LOCUS       sequence-492441-        8110 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Expresses H4-Delta-1-19-Halo under EF1 promoter and can be randomly 
            integrated by PiggyBac transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8110)
  AUTHORS   Semeigazin A, Iida S, Minami K, Tamura S, Ide S, Higashi K, Toyoda 
            A, Kurokawa K, Maeshima K
  TITLE     Behaviors of nucleosomes with mutant histone H4s in euchromatic 
            domains of living human cells.
  JOURNAL   Histochem Cell Biol. 2024 Jul;162(1-2):23-40. doi: 
            10.1007/s00418-024-02293-x. Epub 2024 May 14.
  PUBMED    38743310
REFERENCE   2  (bases 1 to 8110)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Histochem 
            Cell Biol."; date: "2024-07"; volume: "162(1-2):23-40. doi"; pages: 
            " 10.1007/s00418-024-02293-x. Epub 2024 May 14"
COMMENT     Sequence Label: PB-EF1-H4-Delta-1-19-HaloTag-IRES-Neo
FEATURES             Location/Qualifiers
     source          1..8110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             970..1238
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(1251..1462)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   1915..1977
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2107..2124)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2107..2123)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2116..2138)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2265..2720
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2352..2371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2562..2583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2746..2850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2851..2919,2920..3711)
                     /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     complement(3069..3088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3882..4470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4371..4390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4624..4641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4758..4779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4794..4799,4800..4817,4818..4824)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4832..4848
                     /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)."
     primer_bind     4837..4859
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4856..4872
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4856..4872
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5363..5397
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    6160..6281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6171..6190)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6225..6244
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6323..6343
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6403..7197)
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6514..6533)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7124..7143
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_feature    join(7210..7218,7219..7221,7222..7796)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7383..7402)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     7612..7629
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(7834..8110,1..608))
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
ORIGIN
        1 aagcgccaca gtggctcgcg gtcaacagga ttcaggaacg gctcgcggta atggtccatc
       61 tcgacttcag tcagcgggcg gacgacaccc atcggcagcg taccctcgat aaaaacgttc
      121 tgatcgatga tcagcttgcg gccgacgtcg gtggtgcgga aggcctggaa ggtctcgcgg
      181 gcaaattctg gccattcgtc ccaggtcggg atagggcgga tgaactccat aaatgcaata
      241 cctttgacgc gctctggatt gcgcttggcc cagtggaaac ccagagcgga gccccagtcg
      301 tgaatgacca ggacgacctc ttccagaccc agggcttcga tgaaggcatc catgaagcgg
      361 acgtggtcgt cgaagaaata acccaggtct ggtttgtcgg atttgcccat accgatcagg
      421 tctggagcaa tgcagcgatg ggtcggtgca acatgcggga tgatgttgcg ccacacgtag
      481 gaggaggtcg ggttaccgtg caggaacagc acaggggtgc catcgcgcgg accaacatcg
      541 acgtagtgca tgcgctcgcc caggacttcc acataatggg ggtcgaatgg aaagccagta
      601 ccgatttcgg aaccgccacc agagcctcca cctccacttc cacctccgcc tgaaccacct
      661 ccaccagatg gatcaccgcc aaagccataa agggtgcgtc cctggcgctt aagcgcgtag
      721 accacgtcca tggcagtgac tgtcttgcgc ttggcgtgct ccgtataggt gacagcgtca
      781 cggatcacgt tctccaaaaa caccttgagc accccgcgag tctcctcgta gatcagacca
      841 gagatccgct tcacaccgcc acgccgggcc agacgccgga tggccggctt ggtgatgccc
      901 tggatgttgt cacgcaacac cttcatctcg aaattcgcta gctctagagt aggcgccggt
      961 cacagcttgg atctgtaacg gcgcagaaca gaaaacgaaa caaagacgta gagttgagca
     1021 agcagggtca ggcaaagcgt ggagagccgg ctgagtctag gtaggctcca agggagcgcc
     1081 ggacaaaggc ccggtctcga cctgagcttt aaacttacct agacggcgga cgcagttcag
     1141 gaggcaccac aggcgggagg cggcagaacg cgactcaacc ggcgtggatg gcggcctcag
     1201 gtagggcggc gggcgcgtga aggagagatg cgagcccctc gaagcttcag ctgtgttctg
     1261 gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct
     1321 cccccaccct cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg
     1381 cgccaccttc tctaggcacc cgttcaattg ccgacccctc cccccaactt ctcggggact
     1441 gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc gatcgcagat ccttggccgt
     1501 ggaggccttt tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt
     1561 tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg
     1621 gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc
     1681 ccctagcggg ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctca
     1741 taaaagtttt gttactttat agaagaaatt ttgagttttt gtttttttta ataaataaat
     1801 aaacataaat aaattgtttg ttgaatttat tattagtatg taagtgtaaa tataataaaa
     1861 cttaatatct attcaaatta ataaataaac ctcgatatac agaccgataa aacacatgcg
     1921 tcaattttac acatgattat ctttaacgta cgtcacaata tgattatctt tctagggtta
     1981 atctagctgc gtgttctgca gcgtgtcgag catcttcatc tgctccatca cgctgtaaaa
     2041 cacatttgca ccgcgagtct gcccgtcctc cacgggttca aaaacgtgaa tgaacgaggc
     2101 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2161 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2221 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     2281 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2341 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2401 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2461 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2521 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2581 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2641 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2701 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2761 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2821 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2881 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2941 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3001 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3061 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3121 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3181 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3241 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3301 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3361 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3421 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3481 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3541 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3601 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3661 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3721 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3781 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3841 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3901 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3961 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4021 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4081 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4141 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4201 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4261 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4321 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4381 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4441 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4501 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4561 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4621 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4681 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4741 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     4801 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4861 aacagctatg accatgatta cgccaagcgc gcccgccggg taactcacgg ggtatccatg
     4921 tccatttctg cggcatccag ccaggatacc cgtcctcgct gacgtaatat cccagcgccg
     4981 caccgctgtc attaatctgc acaccggcac ggcagttccg gctgtcgccg gtattgttcg
     5041 ggttgctgat gcgcttcggg ctgaccatcc ggaactgtgt ccggaaaagc cgcgacgaac
     5101 tggtatccca ggtggcctga acgaacagtt caccgttaaa ggcgtgcatg gccacacctt
     5161 cccgaatcat catggtaaac gtgcgttttc gctcaacgtc aatgcagcag cagtcatcct
     5221 cggcaaactc tttccatgcc gcttcaacct cgcgggaaaa ggcacgggct tcttcctccc
     5281 cgatgcccag atagcgccag cttgggcgat gactgagccg gaaaaaagac ccgacgatat
     5341 gatcctgatg cagctagatt aaccctagaa agatagtctg cgtaaaattg acgcatgcat
     5401 tcttgaaata ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca
     5461 tctcagtcgc cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga
     5521 ttttgaacta taacgaccgc gtgagtcaaa atgacgcatg attatctttt acgtgacttt
     5581 taagatttaa ctcatacgat aattatattg ttatttcatg ttctacttac gtgataactt
     5641 attatatata tattttcttg ttatagatat cagggacagc ccccccccaa agcccccagg
     5701 gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc
     5761 ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg
     5821 gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac
     5881 acctgggggg atacggggaa aaagccatac caatgggccc taaaaaagga atccagtcaa
     5941 ttccggggct aaacctggct gccactgttt ctttagggac ttcgttcctg tgaggacagg
     6001 aaaccctgca acctggttga ccagagcaag ggatataaga acccagtttg ggaacactgc
     6061 tgtttaaata ttaaacaggg atgtcttgtc ccccggaagg gagaccagat gagcttctga
     6121 tagactaatt gacatcattt gagtcaattc cagacatgat aagatacatt gatgagtttg
     6181 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     6241 ttgctttatt tgtaaccatt ataagctgca ataaacaagt ttagttaacg catgatacaa
     6301 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     6361 caatctttca caaattttgt aatccagagg ttgattgtcg actcagaaga actcgtcaag
     6421 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     6481 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     6541 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     6601 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     6661 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     6721 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     6781 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     6841 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     6901 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     6961 agctgcgcaa ggaacgcccg tcgcggccag ccacgatagc cgcgctgcct cgtcctgcag
     7021 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     7081 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     7141 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcattcc
     7201 ggacgccatg gttgtggcca tattatcatc gtgtttttca aaggaaaacc acgtccccgt
     7261 ggttcggggg gcctagacgt ttttttaacc tcgactaaac acatgtaaag catgtgcacc
     7321 gaggccccag atcagatccc atacaatggg gtaccttctg ggcatccttc agccccttgt
     7381 tgaatacgct tgaggagagc catttgactc tttccacaac tatccaactc acaacgtggc
     7441 actggggttg tgccgccttt gcaggtgtat cttatacacg tggcttttgg ccgcagaggc
     7501 acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt cgctacagac gttgtttgtc
     7561 ttcaagaagc ttccagagga actgcttcct tcacgacatt caacagacct tgcattcctt
     7621 tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa gacagggcca ggtttccggg
     7681 ccctcacatt gccaaaagac ggcaatatgg tggaaaataa catatagaca aacgcacacc
     7741 ggccttattc caagcggctt cggccagtaa cgttaggggg gggggaggga gaggggcgcg
     7801 gccgcggatc cgatttaaat tcgaattaac ttaaccggaa atctccagag tagacagcca
     7861 gcgcgcgatc tcgctgccga tcaggtccgg gttgtcttct tgcagcagat tcagacccgg
     7921 gccgatgtcc acagccttgc agttaggcag gcttttggcc aggcgagcgg cttcggccgg
     7981 tgggatcaga acgcctgggg tgccccagaa cagcagcttc gggacagggg actggtgcag
     8041 ccagtccatg tattcttcga ccagcgcgac gatgttcgct ggctcaccgg cgattggcag
     8101 ctcgtttggg
//
