LOCUS       sequence-37927-e        3679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3679)
  AUTHORS   White MD, Milne RV, Nolan MF
  TITLE     A Molecular Toolbox for Rapid Generation of Viral Vectors to Up- or 
            Down-Regulate Neuronal Gene Expression in vivo.
  JOURNAL   Front Mol Neurosci. 2011;4:8. Epub 2011 Jul 4.
  PUBMED    21772812
REFERENCE   2  (bases 1 to 3679)
  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: "Front Mol 
            Neurosci."; date: "2011"; volume: "4"; pages: "8. Epub 2011 Jul 4"
COMMENT     Sequence Label: pENTR-L3-HCN1miR-EGFP-L2
FEATURES             Location/Qualifiers
     source          1..3679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..138)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(120..138)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(143..159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(143..159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(261..280)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             272..1081
                     /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(339..358)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1228..1816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1717..1736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1970..1987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2146..2173)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2265..2351)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2400..2422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2414..2431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2415..2431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2448..2543
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           2685..2711
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           2775..2816
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     primer_bind     2891..2910
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2942..2951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2948..2950,2951..2953,2954..3667)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2948..2950,2951..2953,2954..3664)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2993..3014)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3254..3273)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3601..3622
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(join(3669..3679,1..89))
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
       61 actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc gcgatatccc
      121 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
      181 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
      241 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
      301 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
      361 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
      421 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
      481 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
      541 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
      601 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
      661 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
      721 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
      781 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
      841 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
      901 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
      961 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     1021 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     1081 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     1141 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     1201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1861 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1921 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1981 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2041 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2101 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     2161 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     2221 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     2281 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
     2341 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc
     2401 ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt
     2461 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat
     2521 gccaactttg tataataaag ttgcttaagg tagccttgca gaagttggtc gtgaggcact
     2581 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     2641 tcgagacaga gaagactctt gcgtttcggt cgacttctag agctctggag gcttgctgaa
     2701 ggctgtatgc tgtaaagacg acagtaggta tcagttttgg ccactgactg actgatacct
     2761 tgtcgtcttt acaggacaca aggcctgtta ctagcactca catggaacaa atggccgaat
     2821 tcggcgctag ctgctgatat cgcatacgcg tggaccagat aggcacctat tggtcttact
     2881 gacatccact ttgcctttct ctccacaggt gtcggtaccg cgggcccggg atccaccggt
     2941 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3001 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3061 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3121 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3181 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3241 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3301 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3361 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3421 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3481 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3541 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3601 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3661 caagtaatac ccagctttc
//
