LOCUS       sequence-172896-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H2B fused to mCherry for chromatin visualization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Nam HS, Benezra R
  TITLE     High levels of Id1 expression define B1 type adult neural stem 
            cells.
  JOURNAL   Cell Stem Cell. 2009 Nov 6. 5(5):515-26.
  PUBMED    19896442
REFERENCE   2  (bases 1 to 6476)
  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: "Cell Stem 
            Cell. 2009 Nov 6. 5(5):515-26."
COMMENT     Sequence Label: H2B-mCherry
FEATURES             Location/Qualifiers
     source          1..6476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(242..670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(352..373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     564..583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    716..940
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     929..946
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        966..984
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     966..983
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(1000..1710)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1278..1297)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1547..1565
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1707..1716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1729..2106)
                     /codon_start=1
                     /gene="HIST1H2BB"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSSK"
     primer_bind     complement(2131..2150)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2132..2150)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2195..2398)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2224..2244)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2399..2778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2950..2969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3044..3148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3149..3217,3218..4009)
                     /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(3367..3386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4180..4768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4669..4688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4922..4939
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5056..5077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5092..5097,5098..5115,5116..5122)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5130..5146
                     /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     5135..5157
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5154..5170
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5154..5170
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5219..5340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5230..5249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5284..5303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5514..6308)
                     /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(5625..5644)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6235..6254
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6375..6476,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6389..6476,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6443..6462)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
      241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
      361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
      421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
      481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
      541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
      601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
      661 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
      721 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
      781 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
      841 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
      901 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgagct
      961 ctagcattta ggtgacacta tagaataggg ccctctagat tacttgtaca gctcgtccat
     1021 gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc
     1081 gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg
     1141 cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt
     1201 cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga
     1261 ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc
     1321 gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt
     1381 caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga
     1441 cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat
     1501 gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag
     1561 cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc
     1621 gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc
     1681 catgttatcc tcctcgccct tgctcaccat ggtggcgacc ggtggatcct tagagctagt
     1741 gtacttggta actgccttag tgccctcgga cacagcatgc ttagccagct ccccaggcag
     1801 cagcaggcgc acagccgtct gaatctccct ggaggtgatg gtcgagcgct tattgtagtg
     1861 agccaggcga gaagcctcgc ccgcgatgcg ctcgaagatg tcgttgacga aggagttcat
     1921 gatccccatg gccttggatg agatgccggt gtcggggtgg acctgcttca gaaccttgta
     1981 cacatagata gaatagctct ccttgcggct gcgcttacgc ttcttaccat ccttcttctg
     2041 cgccttagtg atagccttct tagaaccctt tttaggggct ggagcagact tagagggttc
     2101 aggcatggtg gcggtaccaa gcttgggtct ccctatagtg agtcgtatta atttcgataa
     2161 gccagtaagc agtgggttct ctagttagcc agagagctct gcttatatag acctcccacc
     2221 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     2281 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     2341 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     2401 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     2461 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     2521 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     2581 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     2641 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     2701 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     2761 tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac
     2821 gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt
     2881 aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc
     2941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc
     3001 tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3061 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3121 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3181 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3241 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3301 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3361 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3421 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3481 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3541 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3601 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3661 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3841 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4321 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4801 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4861 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4921 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4981 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5041 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5101 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5161 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
     5221 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     5281 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     5341 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     5401 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     5461 tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag
     5521 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     5581 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     5641 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     5701 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     5761 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     5821 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     5881 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     5941 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     6001 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     6061 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     6121 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     6181 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     6241 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     6301 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atccgaaaat ggatatacaa
     6361 gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     6421 aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat tagtca
//
