LOCUS       sequence-494411-        3827 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  Gateway middle entry clone containing H2B-mScarlet3-S2; Nuclear red 
            fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3827)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 3827)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: pME-H2B-mScarlet3-S2 (JDW 1514)
FEATURES             Location/Qualifiers
     source          1..3827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             687..1064
                     /codon_start=1
                     /gene="HIST1H2BB"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSSK"
     CDS             1080..1121
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1209..1895
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFQYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     1607..1626
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1952..2051)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2068..2087)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2069..2087)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2092..2108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2092..2108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2210..2229)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2221..3030
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2288..2307)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3177..3765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3666..3685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctac tagtgctagc gccaccatgc ctgaaccctc taagtctgct ccagccccta
      721 aaaagggttc taagaaggct atcactaagg cgcagaagaa ggatggtaag aagcgtaagc
      781 gcagccgcaa ggagagctat tctatctatg tgtacaaggt tctgaagcag gtccaccccg
      841 acaccggcat ctcatccaag gccatgggga tcatgaactc cttcgtcaac gacatcttcg
      901 agcgcatcgc gggcgaggct tctcgcctgg ctcactacaa taagcgctcg accatcacct
      961 ccagggagat tcagacggct gtgcgcctgc tgctgcctgg ggagctggct aagcatgctg
     1021 tgtccgaggg cactaaggca gttaccaagt acactagctc taagggtggt gctggtggtg
     1081 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gggatccacc ggtgaattcg
     1141 atggtggagg cggttcaggc ggaggtggct ctggcggtgg cggatcgcca tcaggtacca
     1201 tggtgagcat ggatagcacc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca
     1261 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1321 acgagggcac ccagaccgcc aagctgaggg tgaccaaggg tggccccctg cccttctcct
     1381 gggacatcct gtcccctcag ttccagtacg gctccagggc cttcacgaag caccccgccg
     1441 acatccccga ctactggaag cagtccttcc ccgagggctt caagtgggag cgcgtgatga
     1501 acttcgagga cggcggcgcc gtgtccgtgg cccaggacac ctccctggag gacggcaccc
     1561 tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga
     1621 agaagacaat gggctgggaa gcatccaccg agcggttgta ccccgaggac gtcgtgctga
     1681 agggcgacat taagcatgcc ctgcgcctga aggacggcgg ccggtacctg gcggacttca
     1741 agaccaccta cagggccaag aagcccgtgc agatgcccgg cgccttcaac atcgaccgca
     1801 agttggacat cacatcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg
     1861 tggcccgcca ctccaccggc ggctccggtg gctcctaact ataatgaggc gcgccgtcga
     1921 cgttaacacg cgttaatgat acgtatctag aacccagctt tcttgtacaa agttggcatt
     1981 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     2041 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     2101 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2161 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2221 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2281 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2341 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2401 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2461 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2521 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2581 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2641 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2701 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2761 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2821 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2881 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2941 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3001 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3061 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3121 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3301 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
