LOCUS       (263-bp-DIAL-YB_        3552 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 3552)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3552
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    23..77
                     /label=ZF43/ZF37_hybridpromoter
     promoter        24..48
                     /label=YB_TATA minimal promoter
     regulatory      83..92
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(89..91,92..94,95..808)
                     /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    820..1044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1067..1090
                     /label=pENTR2B.Seq.REV
     CDS             1166..1975
                     /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    2049..2668
                     /label=pBR322_Origin
     rep_origin      2065..2653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      2818..2904
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2996..3023
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    3100..3202
                     /label=ZF43/ZF37_hybridpromoter
     protein_bind    3112..3120
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3116..3124
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3127..3135
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3131..3139
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3142..3150
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3146..3154
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3157..3165
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3161..3169
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3172..3180
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3176..3184
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3187..3195
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3191..3199
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     polyA_signal    3206..3213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3222..3255)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3319..3397
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3519..3552)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cggatttgat ctgatcaagc aactctagag ggtatataat gggggccact agtctactac
       61 cagagctcat cgctagcaat gagccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      121 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtccg
      181 cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac
      241 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttaggctacg gcgtggcctg
      301 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      361 aggctacgtc caggagcgca ccatctcttt caaggacgac ggtacctaca agacccgcgc
      421 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gtgctgaagg gcatcgactt
      481 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaaggt
      541 ctatatcacg gccgacaagc agaagaacgg catcaaggct aacttcaaga cccgccacaa
      601 cgttgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
      661 cggccccgtg ctgctgcccg acaaccacta cctgagccat cagtccaaac tgagcaaaga
      721 ccccaacgag aagcgcgatc acatggtcct gaaggagagg gtgaccgccg ccgggattac
      781 acatgacatg gacgagctgt acaagtaaca acgttgcacc tgtgccttct agttgccagc
      841 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
      901 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
      961 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1021 ctggggatgc ggtgggctct atggcgctcg ctgactgcag gtgcccgtgt ctcaaaatct
     1081 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1141 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     1201 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1261 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     1321 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1381 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctggtgatgc
     1441 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     1501 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     1561 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctagctc aggcgcaatc
     1621 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     1681 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     1741 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1801 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1861 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     1921 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     1981 attggttaat tggttgtaac attattcaga ttgggccccg ttccactgag cgtcagaccc
     2041 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2101 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2161 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2221 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2281 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2341 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2401 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2461 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2521 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2581 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2641 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2701 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2761 tagcatggat ctcggggacg tctaactact aagcgagagt agggaactgc caggcatcaa
     2821 ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2881 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg
     2941 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag
     3001 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttcctgttag ttagttactt
     3061 aagctcgggc ccacctgcag tctacttact cggatcggga gatctgaatt ggagtgagga
     3121 cgtgtagagt gaggacgtgc ggagtgagga cgtgtagagt gaggacgtgc ggagtgagga
     3181 cgtgtagagt gaggacgtgc cctcctctgg atcaaatccg aataacttcg tatagcatac
     3241 attatacgaa gttatctcga gtcgctcggt acgatttaaa ttgaattctg cagtcgacgg
     3301 taccgcgggc ccgggatcct gatagtgacc tgttcgttgc aacaaattga taagcaatgc
     3361 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac caattcagtc
     3421 gactggatcc ggtacctcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     3481 cattttggca aagaattgat taattcgagc gaacgcgtat aacttcgtat agcatacatt
     3541 atacgaagtt at
//
