LOCUS       sequence-168259-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of nfsB-CFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  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 4520)
  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..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        692..710
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     692..709
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      757..766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1423..1432
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1429..1431,1432..1434,1435..2148)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1474..1495)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1735..1754)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2082..2103
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2234..2257
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2401..2535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2425..2444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2479..2498
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(2645..2744)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2761..2780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2762..2780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2785..2801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2785..2801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2903..2922)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2914..3723
                     /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(2981..3000)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3870..4458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4359..4378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc aggctgcagg ctgcaagctt gatttaggtg acactataga atacaagcta
      721 cttgttcttt ttgcaggatc ccatcgattc gaattcgcca ccatggatat catttctgtc
      781 gccttaaagc gtcattccac taaggcattt gatgccagca aaaaacttac cccggaacag
      841 gccgagcaga tcaaaacgct actgcaatac agcccatcca gcaccaactc ccagccgtgg
      901 cattttattg ttgccagcac ggaagaaggt aaagcgcgtg ttgccaaatc cgctgccggt
      961 aattacgtgt tcaacgagcg taaaatgctt gatgcctcgc acgtcgtggt gttctgtgca
     1021 aaaaccgcga tggacgatgt ctggctgaag ctggttgttg accaggaaga tgccgatggc
     1081 cgctttgcca cgccggaagc gaaagccgcg aacgataaag gtcgcaagtt cttcgctgat
     1141 atgcaccgta aagatctgca tgatgatgca gagtggatgg caaaacaggt ttatctcaac
     1201 gtcggtaact tcctgctcgg cgtggcggct ctgggtctgg acgcggtacc catcgaaggt
     1261 tttgacgccg ccatcctcga tgcagaattt ggtctgaaag agaaaggcta caccagtctg
     1321 gtggttgttc cggtaggtca tcacagcgtt gaagatttta acgctacgct gccgaaatct
     1381 cgtctgccgc aaaacatcac cttaaccgaa gtgataccgg tcgccaccat ggtgagcaag
     1441 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1501 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1561 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1621 ctgacctggg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1681 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1741 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1801 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1861 aactacatca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     1921 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1981 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2041 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2101 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgctac
     2161 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     2221 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2281 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     2341 ggcaaaaaga aaaagaagaa atccaaaaca aaatgtgcat taatgtgaag agcggccgta
     2401 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2461 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2521 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2581 gtgtgggagg ttttttaatt cgcggccggc cgcggcgcca atgcattggg cccggtaccc
     2641 gggtacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2701 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2761 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2821 caaaatctct gatgttacat tgcacaagat aaaataatat catcatgaac aataaaactg
     2881 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2941 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     3001 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     3061 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     3121 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     3181 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     3241 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     3301 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     3361 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     3421 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     3481 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     3541 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3601 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3661 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3721 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3781 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4441 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4501 tggccttttg ctcacatgtt
//
