LOCUS       sequence-385191-        3306 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 3306)
  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 3306)
  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: 765 pME nls mCerulean
FEATURES             Location/Qualifiers
     source          1..3306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             144..164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(165..167,168..170,171..881)
                     /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(210..231)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(471..490)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     818..839
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(895..994)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1011..1030)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1012..1030)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1035..1051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1035..1051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1153..1172)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1164..1973
                     /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(1231..1250)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2120..2708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2609..2628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2862..2879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3038..3065)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3157..3243)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3292..3306,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3306,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctggaccatg gctccaaaga agaagcgtaa ggtaatggtg agcaagggcg
      181 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      241 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      301 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      361 cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      421 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      481 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      541 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacg
      601 ccatcagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
      661 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      721 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      781 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      841 ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggggcgcgcc tgacacccag
      901 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
      961 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     1021 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     1081 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1141 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1201 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1261 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1321 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1381 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1441 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     1501 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1561 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     1621 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     1681 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     1741 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     1801 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     1861 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     1921 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     1981 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     2041 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     2101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2761 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2821 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2881 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     2941 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
     3001 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     3061 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     3121 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     3181 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     3241 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
     3301 gacgtt
//
