LOCUS       sequence-385878-        3339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pME fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3339)
  AUTHORS   Kemmler CL, Moran HR, Murray BF, Scoresby A, Klem JR, Eckert RL, 
            Lepovsky E, Bertho S, Nieuwenhuize S, Burger S, D'Agati G, Betz C, 
            Puller AC, Felker A, Ditrychova K, Botschi S, Affolter M, Rohner N, 
            Lovely CB, Kwan KM, Burger A, Mosimann C
  TITLE     Next-generation plasmids for transgenesis in zebrafish and beyond.
  JOURNAL   Development. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531.
  PUBMED    36975217
REFERENCE   2  (bases 1 to 3339)
  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: 
            "Development. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531."
COMMENT     Sequence Label: 769 pME mCerulean-CAAX
FEATURES             Location/Qualifiers
     source          1..3339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..41
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     33..50
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     34..50
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    66..165
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(171..173,174..176,177..887)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(216..237)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(477..496)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     824..845
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(961..1060)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1077..1096)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1078..1096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1101..1117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1101..1117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1219..1238)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1230..2039
                     /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(1297..1316)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2186..2774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2675..2694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2928..2945
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3104..3131)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3223..3309)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
       61 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      121 agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctggacc atggtgagca
      181 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      241 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      301 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      361 ccctgacctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      421 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      481 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      541 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      601 acaacgccat cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
      661 ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      721 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
      781 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      841 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggg cgcgccaagc
      901 tgaaccctcc tgatgagagt ggccccggct gcatgagctg caagtgtgtg ctctcctgac
      961 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1021 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     1081 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1141 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1201 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     1261 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1321 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     1381 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1441 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1501 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     1561 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     1621 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     1681 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     1741 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     1801 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1861 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     1921 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1981 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2041 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2101 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2161 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2221 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2281 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2341 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2401 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2461 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2521 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2581 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2641 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2701 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2761 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2821 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2881 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2941 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3001 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3061 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     3121 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     3181 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     3241 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     3301 ttttatttga tgcctggcag ttccctactc tcgcgttaa
//
