LOCUS       sequence-385843-        3449 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 3449)
  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 3449)
  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: pSN001 pME minprom_mCerulean
FEATURES             Location/Qualifiers
     source          1..3449
                     /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"
     regulatory      296..305
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(302..304,305..307,308..820)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(347..368)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(608..627)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     955..976
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1038..1137)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1154..1173)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1155..1173)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1178..1194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1178..1194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1296..1315)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1307..2116
                     /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(1374..1393)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2263..2851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2752..2771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3005..3022
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3181..3208)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3300..3386)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3435..3449,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3449,1..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 ctccgcggcc gcccccttag atctcccttc caatctgctc agagaggaca
      181 gagtgggcag gagccagcat tgggtatata aagctgagca gggtcagttg cttcttacgt
      241 ttgcttctga gtctgttgtg ttgacttgca acctcagaaa cagacatccg gatccgccac
      301 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      361 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      421 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      481 cctcgtgacc accctgacct ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa
      541 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      601 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      661 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      721 caagctggag tacaacgcca tcagcgacaa cgtctatatc accgccgaca agcagaagaa
      781 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      841 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      901 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
      961 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tggccctcga
     1021 gaagggtggg cgcgccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1081 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1141 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1201 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1261 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1321 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1381 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     1441 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1501 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1561 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     1621 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1681 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1741 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1801 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     1861 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1921 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1981 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2041 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2101 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2161 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2881 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2941 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3001 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3061 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3121 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     3181 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     3241 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     3301 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3361 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     3421 agcatggatg ttttcccagt cacgacgtt
//
