LOCUS       sequence-486048-        3715 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Gateway middle entry clone containing H2B-mCardinal; Nuclear far red
            fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3715)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda J, Ortiz-Guzman J, Turner WD, 
            Largoza G, Sherman M, 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 with diverse applications in vertebrate model 
            systems.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267.
  PUBMED    39026881
REFERENCE   2  (bases 1 to 3715)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267."
COMMENT     Sequence Label: pME-H2B-mCardinal (JDW 1418)
FEATURES             Location/Qualifiers
     source          1..3715
                     /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             694..1071
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1084..1093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1090..1824
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     protein_bind    complement(1840..1939)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1956..1975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1957..1975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1980..1996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1980..1996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2098..2117)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2109..2918
                     /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(2176..2195)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3065..3653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3554..3573
                     /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 agcaggcagc cggtggatcc agccaccgcc accatgccag agccagcgaa gtctgctccc
      721 gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag
      781 cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc
      841 caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac
      901 attttcgagc gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc
      961 atcacctcca gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag
     1021 cacgccgtgt ccgagggtac taaggccatc accaagtaca ccagcgctaa ggatccaccg
     1081 gtcgccacca tggtgagcaa gggcgaggag ctgatcaagg agaacatgca catgaagctg
     1141 tacatggaag gcaccgtgaa caaccaccac ttcaagtgca ccaccgaagg ggagggcaag
     1201 ccctacgagg gcacccagac ccagaggatt aaggtggtgg agggaggccc cctgccgttc
     1261 gcattcgaca tcctggccac ctgctttatg tacgggagca agaccttcat caaccacacc
     1321 cagggcatcc ccgatttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
     1381 accacatacg aagacggggg cgtgcttacc gttacccagg acaccagcct ccaggacggc
     1441 tgcttgatct acaacgtcaa gctcagaggg gtgaacttcc catccaacgg ccctgtgatg
     1501 cagaagaaaa cactcggctg ggaggccacc accgagaccc tgtaccccgc tgacggcggc
     1561 ctggaaggca gatgcgacat ggccctgaag ctcgtgggcg ggggccacct gcactgcaac
     1621 ctgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
     1681 tttgtggacc gcagactgga aagaatcaag gaggccgaca atgagaccta cgtcgagcag
     1741 cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat
     1801 ggcatggacg agctgtacaa gtaataaacg cgtactagtg cccagctttc ttgtacaaag
     1861 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     1921 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     1981 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2041 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2101 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2161 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2221 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2281 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2341 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2401 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2461 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2521 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     2581 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2641 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     2701 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     2761 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     2821 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     2881 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     2941 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3001 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3061 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3181 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
