LOCUS       Exported                4203 bp ds-DNA     circular SYN 20-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4203)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 10, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4203
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     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"
     regulatory      689..698
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     691..716
                     /label=FWD
     CDS             695..1186
                     /codon_start=1
                     /label=EGFP N-terminus
                     /label=EGFP 5' and kozak
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     V"
     intron          join(1188..1256,1883..2072)
                     /label=Artificial intron and MCS
     intron          1257..1882
                     /label=mmClcn5_intron
                     /label=miR-500
                     /note="bp   6812809 and 6816900, in introns encoding Clcn5,
                     on X chromosome of mouse   "
     ncRNA           1512..1603
                     /label=miR-500
                     /ncRNA_class="miRNA"
     CDS             2073..2299
                     /codon_start=1
                     /label=3' end of EGFP
                     /translation="NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALS
                     KDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(2287..2309)
                     /label=REV
     protein_bind    complement(2326..2425)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2443..2461)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2466..2482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2595..3404
                     /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"
     rep_origin      3553..4141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccgc caccatggtg agcaagggcg aggagctgtt
      721 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      781 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      841 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      901 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      961 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1021 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1081 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1141 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgagtg gatcgggtac
     1201 cgaattccat gatctttttt tttttttttt ttttcccatg atcatcccgg ggatccacca
     1261 tgctgagcag gtggtagtcc ccaagataga ctagctctca ccctcaaaag agatgcgtag
     1321 ggagtggcat ctccatgttt tcagttgatg gtgagtcaca aaatgggact ttttaatcta
     1381 cagaaacaca caaagtgtaa gacttctcat ttctgctttc atattggtgt atagtacagt
     1441 gcttcccgcc ccccccatct cccccccccg cctccccccc tgcacattga gtactcaagg
     1501 ccctggtcct tctcctctgc tccccctctc taatccttgc tatctgggtg cttagtgcta
     1561 tctcaatgca atgcacctgg gcaagggttc agagaaggtg agctcatctg catcaagaag
     1621 gaccagagac cctgtgtgtc agcctacccc tccctccttc tttccccctc ccccttcctt
     1681 ccctctattg agtaaaatag ttgtttgagt gagtcatgta taccagggaa ccccctaaca
     1741 gattatttaa gagatttcag gtactctagt tggcttcttt ctacagctaa cttataaaat
     1801 actcaggtac ctggagtgtg taggctttct ctgtattctg tcctgtatgg ttatacagga
     1861 tgaggtcagg agttcaaacc tgctgcagat gcagtcagat ttctgaactg tctcagcaaa
     1921 tggtaagtaa cttagagcta gtaattaata acctgtcctt tgatttctga ttcagccaag
     1981 aatggccata tttgggaaag gcagatctgg agagtaacca cgttttcatg cagtctagac
     2041 tgttaacgca gtgtttctct aactctcagg tgaacttcaa gatccgccac aacatcgagg
     2101 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2161 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2221 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2281 tggacgagct gtacaagtaa agccgccgca agggtgggcg cgccgaccca gctttcttgt
     2341 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     2401 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     2461 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2521 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     2581 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     2641 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2701 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2761 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2821 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2881 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2941 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     3001 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     3061 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     3121 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     3181 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     3241 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     3301 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     3361 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     3421 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     3481 aatcccttac caagtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4201 gtt
//
