LOCUS       sequence-142265-        7505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression plasmid for human histone H4 with C-terminal 
            BirA-biotinylation signal and TEV celavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7505)
  AUTHORS   Gabriel CH, Gross F, Karl M, Stephanowitz H, Hennig AF, Weber M, 
            Gryzik S, Bachmann I, Hecklau K, Wienands J, Schuchhardt J, Herzel 
            H, Radbruch A, Krause E, Baumgrass R
  TITLE     Identification of Novel Nuclear Factor of Activated T Cells 
            (NFAT)-Associated Proteins in T cells.
  JOURNAL   J Biol Chem. 2016 Sep 16. pii: jbc.M116.739326.
  PUBMED    27637333
REFERENCE   2  (bases 1 to 7505)
  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"; journalName: "J Biol 
            Chem. 2016 Sep 16. pii: jbc.M116.739326."
COMMENT     Sequence Label: pMIG-hH4-AVITEV
FEATURES             Location/Qualifiers
     source          1..7505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             43..560
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    624..965
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1032..1448
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1378..1400
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1416..1432
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1800..1823
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1830..1850
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1863..1907
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     1946..1962
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1947..1963
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1982..2548
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1998..2550
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2142..2159)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2369..2388
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2562..2564,2565..2567,2568..3281)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2562..2564,2565..2567,2568..3278)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2607..2628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2868..2887)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3215..3236
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3372..3887
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4056..4072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4056..4072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4069..4091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4080..4096
                     /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(4104..4110,4111..4128,4129..4134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4149..4170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4287..4304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4458..5046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4538..4557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5217..6008,6009..6077))
                     /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     5840..5859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6078..6182)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6250..6268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6306..6328)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6428..6447
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6470..6490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6475..6804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6655..6790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6717..6736
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6852..6899
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     7093..7115
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7107..7124
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7108..7124
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7327..7346
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
       61 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
      121 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
      181 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
      241 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
      301 acctgaaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
      361 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
      421 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
      481 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
      541 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcctagggac
      601 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
      661 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
      721 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      781 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      841 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
      901 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
      961 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1021 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1081 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1141 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1201 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1261 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1321 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1381 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1441 taggcgccgg aattagatct gcggccgcgg atttaaatcc caccatgagc ggcagaggca
     1501 agggcggcaa gggactggga aaaggcggag ccaagcggca ccggaaggtg ctgagagaca
     1561 acatccaggg catcaccaag cccgccatca gacggctggc tagaagaggc ggcgtgaaga
     1621 gaatcagcgg cctgatctac gaagagacac ggggcgtgct gaaggtgttc ctggaaaacg
     1681 tgatccggga cgccgtgacc tacaccgagc acgccaagag aaagaccgtg accgccatgg
     1741 acgtggtgta cgccctgaag agacagggca gaaccctgta cggcttcggc ggctacgtag
     1801 attacaagga tgacgacgat aagcttgtag agaacctgta cttccagggc gttaacggcg
     1861 gtggcctgaa cgatatcttc gaggcacaga aaatcgaatg gcatgaggtt aactgagcga
     1921 tcgcttataa gcggccgcgt cgaggtcgag gtcgacggta tcgataagct tgatatcgaa
     1981 ttccgccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     2041 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2101 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     2161 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     2221 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2281 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     2341 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     2401 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     2461 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2521 ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg
     2581 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2641 agttcagcgt gtctggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2701 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2761 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2821 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2881 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2941 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3001 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3061 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3121 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3181 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3241 ccgccgggat cactctcggc atggacgagc tgtacaagta agaattcctg cagcccgggg
     3301 gatccgtcga cctgcagcca agcttatcga taaaataaaa gattttattt agtctccaga
     3361 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa ttaacgccat
     3421 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     3481 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     3541 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     3601 catcagatgt ttccagggtg ccccaaggac ctgaaaatga ccctgtgcct tatttgaact
     3661 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     3721 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
     3781 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     3841 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc
     3901 ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa
     3961 ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc
     4021 gcagaagagc tggggagaat taattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4081 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4141 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4261 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4321 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4381 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4441 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4501 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4561 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4621 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4681 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4741 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4801 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4861 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4921 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4981 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5041 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5101 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5161 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5221 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5281 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5341 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5401 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5461 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5521 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5581 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5641 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5701 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5761 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5821 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5881 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5941 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6001 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6061 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6121 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6181 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6241 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6301 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6361 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6421 aactatgcgg catcagagca gattgtactg agagtgcacc atatgctgtg gaatgtgtgt
     6481 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6541 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6601 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6661 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6721 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     6781 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     6841 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     6901 aggtgaggaa ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6961 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     7021 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     7081 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctcgctctcc
     7141 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     7201 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     7261 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     7321 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     7381 ggccacgatg cgtccggcgt agaggatcga ttagtccaat ttgttaaaga caggatatca
     7441 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     7501 ccata
//
