LOCUS       sequence-174929-        4568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  membrane targeted EYFP expression in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4568)
  AUTHORS   Yoshimatsu T, D'Orazi FD, Gamlin CR, Suzuki SC, Suli A, Kimelman D, 
            Raible DW, Wong RO
  TITLE     Presynaptic partner selection during retinal circuit reassembly 
            varies with timing of neuronal regeneration in vivo.
  JOURNAL   Nat Commun. 2016 Feb 3;7:10590. doi: 10.1038/ncomms10590.
  PUBMED    26838932
REFERENCE   2  (bases 1 to 4568)
  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: "Nat 
            Commun."; date: "2016-02-3"; volume: "7:10590. doi"; pages: " 
            10.1038/ncomms10590"
COMMENT     Sequence Label: pUAS-MEYFP
FEATURES             Location/Qualifiers
     source          1..4568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      468..477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(474..476,477..479,480..1193)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(519..540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(780..799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1127..1148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1316..1437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1353..1372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1407..1426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1444..1899)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1581..1602)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1793..1812
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1926..2030
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2032..2389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2032..2223
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2055..2075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2240..2375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2424..3218
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2424..3218
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2478..2497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3088..3107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3406..3425)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3450..3497
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3826..4414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4315..4334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcttaggcc tccaaggcgg agtactgtcc tccgggctgg cggagtactg tcctccggca
       61 aggtcggagt actgtcctcc gacactagag gtcggagtac tgtcctccga cgcaaggcgg
      121 agtactgtcc tccgggctgc ggagtactgt cctccggcaa ggtcggagta ctgtcctccg
      181 acactagagg tcggagtact gtcctccgac gcaaggtcgg agtactgtcc tccgacgcaa
      241 ggcggagtac tgtcctccgg gctggcggag tactgtcctc cggcaagggt cgactctaga
      301 gggtatataa tggatcccat cgcgtctcag cctcactttg agctcctcca cacgaattca
      361 cagccaccat gctgtgctgc atcagaagaa ctaaaccggt tgagaagaat gaagaggccg
      421 atcaggagct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtga
      481 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      541 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      601 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      661 ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
      721 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      781 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      841 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      901 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
      961 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1021 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1081 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1141 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     1201 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1261 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1321 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1381 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     1441 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1501 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1561 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1621 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1681 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1741 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1801 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1861 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     1921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1981 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2041 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2101 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2161 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2221 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2281 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2341 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     2401 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2461 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2521 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2581 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     2641 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2701 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2761 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2821 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2881 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2941 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3001 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3061 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3121 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3181 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3241 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3301 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3361 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     3421 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3481 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3541 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3601 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3661 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3721 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3781 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3841 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3901 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3961 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4021 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4081 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4141 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4201 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4261 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4321 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4381 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4441 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4501 tgtggataac cgtattaccg ccatgcatta gttattacta gcgctaccgg actcagatct
     4561 cgagctca
//
