LOCUS       sequence-192948-        5188 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            mCherry in the pCS2+ expression vector. It has the SP6 RNA 
            polymerase site for in vitro transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5188)
  AUTHORS   Wan Y, Wei Z, Looger LL, Koyama M, Druckmann S, Keller PJ
  TITLE     Single-Cell Reconstruction of Emerging Population Activity in an 
            Entire Developing Circuit.
  JOURNAL   Cell. 2019 Oct 3;179(2):355-372.e23. doi: 
            10.1016/j.cell.2019.08.039. Epub 2019 Sep 26.
  PUBMED    31564455
REFERENCE   2  (bases 1 to 5188)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2019.08.039"; journalName: "Cell"; date: "2019-10-3-
            3"; volume: "179"; issue: "2"; pages: "355-372.e23"
COMMENT     Sequence Label: pCS2+ H2B-mCherry
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..1043
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1078..1096
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1078..1095
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1147..1524
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1537..1546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1543..2250
                     /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(1688..1706)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1956..1975
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2150..2173
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2283..2417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2307..2326)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2361..2380
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2530..2550)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2530..2548)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2569..2585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2569..2585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2582..2604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2593..2609
                     /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(2617..2623,2624..2641,2642..2647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2662..2683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2800..2817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2971..3559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3051..3070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3730..4521,4522..4590))
                     /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     4353..4372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4591..4695)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4721..5176)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4858..4879)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5070..5089
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt
       61 cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
      121 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
      181 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc
      241 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
      301 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
      361 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
      421 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
      481 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
      541 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
      601 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
      661 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
      721 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
      781 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
      841 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
      901 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
      961 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     1021 taagcaatgc tcgtttaggg aaccgccatt ctgcctgggg acgtcggagc aagcttgatt
     1081 taggtgacac tatagaatac aagctacttg ttctttttgc aggatcccat cgacggtacc
     1141 gccaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
     1201 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
     1261 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
     1321 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     1381 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     1441 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     1501 atcaccaagt acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag
     1561 gaggacaaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     1621 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     1681 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg
     1741 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     1801 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     1861 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     1921 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     1981 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     2041 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ccgaggtcaa gaccacctac
     2101 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gctggacatc
     2161 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgcgccga gggccgccac
     2221 tccaccggcg gcatggacga gctgtacaag ttatctagaa ctatagtgag tcgtattacg
     2281 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2341 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2401 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2461 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     2521 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     2581 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     2641 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2701 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2761 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2821 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3601 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3661 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3721 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3781 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3841 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3901 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3961 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4021 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4081 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4141 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4201 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4261 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4321 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4381 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4441 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4501 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4561 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4621 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4681 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     4741 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4801 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4861 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4921 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4981 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5041 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5101 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5161 ccgctacagg gcgcgtccca ttcgccat
//
