LOCUS       sequence-330064-        5196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H2B fused to pr-mEosFP for chromatin visualization and primed 
            conversion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5196)
  AUTHORS   Welling M, Kalyviotis K, Pantazis P
  TITLE     Primed Track: Reliable Volumetric Single-cell Tracking and Lineage 
            Tracing of Living Specimen with Dual-labeling Approaches.
  JOURNAL   Bio Protoc. 2020 Jun 5;10(11):e3645. doi: 10.21769/BioProtoc.3645. 
            eCollection 2020 Jun 5.
  PUBMED    33659315
REFERENCE   2  (bases 1 to 5196)
  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: "Bio 
            Protoc."; date: "2020-06-5"; volume: "10(11):e3645. doi"; pages: " 
            10.21769/BioProtoc.3645. eCollection 2020 Jun 5"
COMMENT     Sequence Label: pCS2+-H2B-pr-mEosFP
FEATURES             Location/Qualifiers
     source          1..5196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..52
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     34..51
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             157..534
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             553..1233
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFTEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    1247..1381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1271..1290)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1325..1344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1494..1514)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1494..1512)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1533..1549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1533..1549)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1546..1568)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1557..1573
                     /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(1581..1587,1588..1605,1606..1611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1626..1647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1764..1781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1935..2523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2015..2034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2694..3485,3486..3554))
                     /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     3317..3336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3555..3659)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3685..4140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3822..3843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4034..4053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4211..5195
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
       61 tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggcctctcga gcctctagaa
      121 ctatagtgag tcgtattact cgacggtacc gccaccatgc cagagccagc gaagtctgct
      181 cccgccccga aaaagggctc caagaaggcg gtgactaagg cgcagaagaa aggcggcaag
      241 aagcgcaagc gcagccgcaa ggagagctat tccatctatg tgtacaaggt tctgaagcag
      301 gtccaccctg acaccggcat ttcgtccaag gccatgggca tcatgaattc gtttgtgaac
      361 gacattttcg agcgcatcgc aggtgaggct tcccgcctgg cgcattacaa caagcgctcg
      421 accatcacct ccagggagat ccagacggcc gtgcgcctgc tgctgcctgg ggagttggcc
      481 aagcacgccg tgtccgaggg tactaaggcc atcaccaagt acaccagcgc taaggatcca
      541 ccggtcgcca ccatgagtgc gattaagcca gacatgaaga tcaacctccg tatggaaggc
      601 aacgtaaacg ggcaccactt tgtgatcgac ggagatggta caggcaagcc ttttgaggga
      661 aaacagagta tggatcttga agtcaaagag ggcggacctc tgccttttgc ctttgatatc
      721 ctgaccactg cattccatta cggcaacagg gtattcaccg aatatccaga ccacatacaa
      781 gactatttta agcagtcgtt tcctaagggg tattcgtggg aacgaagctt gactttcgaa
      841 gacgggggca tttgcattgc cagaaacgac ataacaatgg aaggggacac tttctataat
      901 aaagttcgat ttcacggtac aaactttccc gccaatggtc cagttatgca gaagaagacg
      961 ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg atggagtgct gacgggtgat
     1021 attcacatgg ctttgttgct tgaaggaaat gcccattacc gatgtgactt cagaactact
     1081 tacaaagcta aggagaaggg tgtcaagtta ccaggctacc actttgtgga ccactgcatt
     1141 gagattttaa gccatgacaa agattacaac aaggttaagc tgtatgagca tgctgttgct
     1201 cattctggat tgcctgacaa tgccagacga taagtcgtat tacgtagatc cagacatgat
     1261 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1321 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1381 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1441 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     1501 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1561 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     1621 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1681 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1741 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1801 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1861 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2101 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2341 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaatg gaataagggc gacacggaaa
     3541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3661 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     3721 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3781 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3841 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3901 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3961 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4021 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4081 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4141 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4201 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     4261 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     4321 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     4381 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     4441 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     4501 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     4561 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     4621 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     4681 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     4741 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     4801 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     4861 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     4921 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     4981 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
     5041 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     5101 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     5161 gggaggtcta tataagcaat gctcgtttag ggaacc
//
