LOCUS       sequence-183638-        8093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses H2B protein tagged with eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  AUTHORS   Sottile F, Aulicino F, Theka I, Cosma MP
  TITLE     Mesenchymal stem cells generate distinct functional hybrids in vitro
            via cell fusion or entosis.
  JOURNAL   Sci Rep. 2016 Nov 9;6:36863. doi: 10.1038/srep36863.
  PUBMED    27827439
REFERENCE   2  (bases 1 to 8093)
  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: "Sci Rep."; 
            date: "2016-11-9"; volume: "6:36863. doi"; pages: " 
            10.1038/srep36863"
COMMENT     Sequence Label: pHIV-H2B-eGFP
FEATURES             Location/Qualifiers
     source          1..8093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        31..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             628..808
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    855..980
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1477..1710
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1895..1939
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2237..2354
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2417..3595
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2648..3586
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3543..3563
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(3641..4215,4216..4218,4219..4227)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3808..3825)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4035..4054
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4228..4608
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MSPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYK
                     VLKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLL
                     PGELAKHAVSEGTKAITKYTSAK"
     regulatory      4621..4630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4627..4629,4630..4632,4633..5346)
                     /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(4627..4629,4630..4632,4633..5343)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4672..4693)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4933..4952)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5280..5301
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5366..5382)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5367..5383)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5892..6072
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6134..6722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6214..6233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6893..7684,7685..7753))
                     /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     7516..7535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7754..7858)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7933..7952)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
      661 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      721 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      781 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      841 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      901 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      961 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1021 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1081 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1141 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1201 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1261 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1321 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1381 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1441 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1501 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1561 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1621 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1681 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1741 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1801 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1861 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1921 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1981 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     2041 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     2101 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     2161 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg
     2221 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     2281 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     2341 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg ttagtaccgg
     2401 gcccgctcta gcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     2461 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
     2521 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2581 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2641 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     2701 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
     2761 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
     2821 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
     2881 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
     2941 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
     3001 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
     3061 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     3121 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     3181 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     3241 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     3301 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     3361 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     3421 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     3481 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     3541 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgagcggc
     3601 cgctgagtta actattctag agtacccggg ctaggatccg cccctctccc tccccccccc
     3661 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     3721 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     3781 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     3841 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     3901 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     3961 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     4021 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     4081 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     4141 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     4201 aaacacgatg ataatatggc cacaaccatg tcaccagagc cagcgaagtc tgctcccgcc
     4261 ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc
     4321 aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac
     4381 cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt
     4441 ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc
     4501 acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac
     4561 gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagga tccaccggtc
     4621 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4681 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4741 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     4801 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     4861 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4921 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4981 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     5041 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     5101 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     5161 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5221 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     5281 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     5341 aagtaaagcg gccgcgacat gacatcgata ccgtcgacct cgatcgagac ctagaaaaac
     5401 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
     5461 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
     5521 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5581 taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
     5641 acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg
     5701 gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag
     5761 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
     5821 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
     5881 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     5941 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6001 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6061 aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6661 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7741 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7801 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7861 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7921 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7981 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     8041 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgc
//
