LOCUS       sequence-174930-        4032 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expression of nfsB-CFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4032)
  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 4032)
  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: pME-nfsB-CFP
FEATURES             Location/Qualifiers
     source          1..4032
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(1..100)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(117..136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(118..136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(141..157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(141..157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(259..278)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             270..1079
                     /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(337..356)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1226..1814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1715..1734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1968..1985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2144..2171)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2263..2349)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2398..2420
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2412..2429
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2413..2429
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2445..2544
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2568..2586
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2568..2585
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2633..2642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3299..3308
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3305..3307,3308..3310,3311..4024)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3350..3371)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3611..3630)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3958..3979
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
       61 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
      121 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
      181 atctctgatg ttacattgca caagataaaa taatatcatc atgaacaata aaactgtctg
      241 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
      301 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
      361 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
      421 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
      481 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
      541 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
      601 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
      661 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
      721 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
      781 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
      841 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
      901 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
      961 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1021 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     1081 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     1141 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     1201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1981 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2041 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2101 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     2161 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     2221 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     2281 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     2341 ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc
     2401 agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat
     2461 tttgactgat agtgacctgt tcgttgcaac acattgatga gcaatgcttt tttataatgc
     2521 caactttgta caaaaaagca ggctgcaggc tgcaggctgc aagcttgatt taggtgacac
     2581 tatagaatac aagctacttg ttctttttgc aggatcccat cgattcgaat tcgccaccat
     2641 ggatatcatt tctgtcgcct taaagcgtca ttccactaag gcatttgatg ccagcaaaaa
     2701 acttaccccg gaacaggccg agcagatcaa aacgctactg caatacagcc catccagcac
     2761 caactcccag ccgtggcatt ttattgttgc cagcacggaa gaaggtaaag cgcgtgttgc
     2821 caaatccgct gccggtaatt acgtgttcaa cgagcgtaaa atgcttgatg cctcgcacgt
     2881 cgtggtgttc tgtgcaaaaa ccgcgatgga cgatgtctgg ctgaagctgg ttgttgacca
     2941 ggaagatgcc gatggccgct ttgccacgcc ggaagcgaaa gccgcgaacg ataaaggtcg
     3001 caagttcttc gctgatatgc accgtaaaga tctgcatgat gatgcagagt ggatggcaaa
     3061 acaggtttat ctcaacgtcg gtaacttcct gctcggcgtg gcggctctgg gtctggacgc
     3121 ggtacccatc gaaggttttg acgccgccat cctcgatgca gaatttggtc tgaaagagaa
     3181 aggctacacc agtctggtgg ttgttccggt aggtcatcac agcgttgaag attttaacgc
     3241 tacgctgccg aaatctcgtc tgccgcaaaa catcacctta accgaagtga taccggtcgc
     3301 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3361 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     3421 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3481 caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat
     3541 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3601 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3661 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3721 gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa
     3781 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3841 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3901 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3961 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4021 gtaaagcggc cg
//
