LOCUS       (nested-DIAL-YB_        3488 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3488)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3488
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    156..179
                     /label=pENTR2B.Seq.REV
     CDS             255..1064
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    1138..1757
                     /label=pBR322_Origin
     rep_origin      1154..1742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      1907..1993
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2085..2112
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    2189..2291
                     /label=ZF43/ZF37_hybridpromoter
     protein_bind    2201..2209
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2205..2213
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2216..2224
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2220..2228
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2231..2239
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2235..2243
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2246..2254
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2250..2258
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2261..2269
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2265..2273
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2276..2284
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2280..2288
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    complement(2311..2344)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2365..2398
                     /label=VloxP
     protein_bind    2399..2424
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    2400..2424
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2490..2523
                     /label=VloxP
     protein_bind    complement(2544..2577)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2600..2654
                     /label=ZF43/ZF37_hybridpromoter
     promoter        2601..2625
                     /label=YB_TATA minimal promoter
     regulatory      2660..2669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2666..2668,2669..2671,2672..3385)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     polyA_signal    join(3397..3488,1..133)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
       61 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
      121 gtgggctcta tggcgctcgc tgactgcagg tgcccgtgtc tcaaaatctc tgatgttaca
      181 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
      241 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
      301 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
      361 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
      421 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
      481 ttatgcctct tccgaccatc aagcatttta tccgtactcc tggtgatgca tggttactca
      541 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
      601 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
      661 attgtccttt taacagcgat cgcgtatttc gtctagctca ggcgcaatca cgaatgaata
      721 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
      781 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
      841 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
      901 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
      961 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1021 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     1081 ggttgtaaca ttattcagat tgggccccgt tccactgagc gtcagacccc gtagaaaaga
     1141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1261 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1801 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgct agcatggatc
     1861 tcggggacgt ctaactacta agcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     1921 ggctcagtcg gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     1981 gagtaggaca aatccgccgg gagcggattt gaacgttgtg aagcaacggc ccggagggtg
     2041 gcgggcagga cgcccgccat aaactgccag gcatcaaact aagcagaagg ccatcctgac
     2101 ggatggcctt tttgcgtttc tacaaactct tcctgttagt tagttactta agctcgggcc
     2161 cacctgcagt ctacttactc ggatcgggag atctgaattg gagtgaggac gtgtagagtg
     2221 aggacgtgcg gagtgaggac gtgtagagtg aggacgtgcg gagtgaggac gtgtagagtg
     2281 aggacgtgcc ctcctctgga tcaaatccga ataacttcgt atagcataca ttatacgaag
     2341 ttatctcgag tcgctcggta cgattcaatt tctgagaact gtcattctcg gaaattgagc
     2401 caactttgta caaaaaagca ggctccacca tgggaaccaa ttcagtcgac tggatccggt
     2461 acctcctaca gctcctgggc aacgtgctgt caatttctga gaactgtcat tctcggaaat
     2521 tgaattaatt cgagcgaacg cgtataactt cgtatagcat acattatacg aagttatcgg
     2581 atttgatctg atcaagcaac tctagagggt atataatggg ggccactagt ctactaccag
     2641 agctcatcgc tagcaatgag ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     2701 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtccgcgg
     2761 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     2821 caagctgccc gtgccctggc ccaccctcgt gaccacctta ggctacggcg tggcctgctt
     2881 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2941 ctacgtccag gagcgcacca tctctttcaa ggacgacggt acctacaaga cccgcgccga
     3001 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgtg ctgaagggca tcgacttcaa
     3061 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaaggtcta
     3121 tatcacggcc gacaagcaga agaacggcat caaggctaac ttcaagaccc gccacaacgt
     3181 tgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3241 ccccgtgctg ctgcccgaca accactacct gagccatcag tccaaactga gcaaagaccc
     3301 caacgagaag cgcgatcaca tggtcctgaa ggagagggtg accgccgccg ggattacaca
     3361 tgacatggac gagctgtaca agtaacaacg ttgcacctgt gccttctagt tgccagccat
     3421 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3481 tttcctaa
//
