LOCUS       sequence-150107-        3699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear mCerulean2 (blue) fluorophore for use with 5' promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3699)
  AUTHORS   Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan 
            A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ
  TITLE     A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous 
            System and Neurodegenerative Disease.
  JOURNAL   Zebrafish. 2016 Sep 15.
  PUBMED    27631880
REFERENCE   2  (bases 1 to 3699)
  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: "Zebrafish. 
            2016 Sep 15."
COMMENT     Sequence Label: pME-H2B-mCerulean3
FEATURES             Location/Qualifiers
     source          1..3699
                     /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"
     primer_bind     complement(1404..1423)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1824..1923)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1940..1959)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1941..1959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1964..1980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1964..1980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2082..2101)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2093..2902
                     /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(2160..2179)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3049..3637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3538..3557
                     /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 agcaggctga attcgccgcc accatgcctg aacctagcaa gagcgcccct gcccctaaga
      721 agggaagcaa gaaggccatc accaaggccc agaagaagga cggaaagaag cgcaagagaa
      781 gccgcaaaga gtcttacagc atctacgtgt acaaggtgct gaaacaggtg caccctgaca
      841 ccggaatcag cagcaaggcc atgggaatca tgaacagctt cgtgaacgac atcttcgaga
      901 gaatcgccgg cgaggccagc agactggccc actacaacaa gagaagcacc atcaccagcc
      961 gcgagatcca gaccgccgtc agactgctgc tgcctggcga gctggccaag cacgccgtgt
     1021 ctgagggaac aaaggccgtg accaagtaca caagcagcaa gggcggagga agcggaggcg
     1081 gaagcggcgg aggatctatg gtgtctaagg gcgaggaact gttcaccggc gtggtgccta
     1141 tcctggtgga actggacggc gacgtgaacg gacacaagtt cagcgtgtcc ggcgagggcg
     1201 aaggcgacgc cacctacgga aagctgaccc tgaagttcat ctgcaccacc ggaaagctgc
     1261 ctgtgccttg gcctaccctg gtcaccaccc tgagctgggg agtgcagtgc ttcgccagat
     1321 accctgacca catgaagcag cacgatttct tcaagagcgc catgcctgag ggatacgtgc
     1381 aggaaagaac catcttcttc aaggacgacg gcaactacaa gaccagagcc gaagtgaagt
     1441 tcgagggcga caccctggtc aacagaatcg agctgaaggg aatcgacttc aaagaggacg
     1501 gaaacatcct gggacacaag ctggagtaca acgccatcca cggaaacgtg tacatcaccg
     1561 ccgacaagca gaagaacgga atcaaggcca acttcggcct gaactgcaac atcgaggacg
     1621 gaagcgtgca gctggccgac cactaccagc agaacacccc tatcggagat ggacctgtgc
     1681 tgctgccaga caaccactac ctgagcaccc agagcaagct gagcaaggac cctaacgaga
     1741 agcgcgacca catggtgctg ctggaatttg tgaccgccgc tggaatcacc ctgggcatgg
     1801 atgaactgta taaatgaact agtacccagc tttcttgtac aaagttggca ttataagaaa
     1861 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1921 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1981 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2041 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2101 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2161 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2221 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2281 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2341 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2401 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2461 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2521 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2581 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2641 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2701 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2761 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2821 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2881 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     2941 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3001 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3661 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
