LOCUS       (27-bp-DIAL-YB_T        3316 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 3316)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    23..46
                     /label=pENTR2B.Seq.REV
     CDS             122..931
                     /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    1005..1624
                     /label=pBR322_Origin
     rep_origin      1021..1609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      1774..1860
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1952..1979
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    2056..2158
                     /label=ZF43/ZF37_hybridpromoter
     protein_bind    2068..2076
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2072..2080
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2083..2091
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2087..2095
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2098..2106
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2102..2110
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2113..2121
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2117..2125
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2128..2136
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2132..2140
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    2143..2151
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2147..2155
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     misc_feature    2162..2169
                     /label=SV40 polyA
     polyA_signal    2162..2169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2178..2211
                     /label=loxP
                     /label=loxP(1)
     protein_bind    2178..2211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2239..2272
                     /label=loxP
     protein_bind    2239..2272
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2295..2349
                     /label=ZF43/ZF37_hybridpromoter
     promoter        2296..2320
                     /label=YB_TATA minimal promoter
     regulatory      2355..2364
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2361..2363,2364..2366,2367..3080)
                     /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    3092..3316
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgctcgctga ctgcaggtgc ccgtgtctca aaatctctga tgttacattg cacaagataa
       61 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
      121 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
      181 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
      241 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
      301 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
      361 gaccatcaag cattttatcc gtactcctgg tgatgcatgg ttactcacca ctgcgatccc
      421 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
      481 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
      541 cagcgatcgc gtatttcgtc tagctcaggc gcaatcacga atgaataacg gtttggttga
      601 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
      661 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
      721 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
      781 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
      841 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
      901 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta
      961 ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1681 gttatcccct gattctgtgg ataaccgtat taccgctagc atggatctcg gggacgtcta
     1741 actactaagc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
     1801 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1861 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
     1921 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
     1981 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
     2041 cttactcgga tcgggagatc tgaattggag tgaggacgtg tagagtgagg acgtgcggag
     2101 tgaggacgtg tagagtgagg acgtgcggag tgaggacgtg tagagtgagg acgtgccctc
     2161 ctctggatca aatccgaata acttcgtata gcatacatta tacgaagtta tctcgagtcg
     2221 ctcggtacgc gaacgcgtat aacttcgtat agcatacatt atacgaagtt atcggatttg
     2281 atctgatcaa gcaactctag agggtatata atgggggcca ctagtctact accagagctc
     2341 atcgctagca atgagccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2401 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg
     2461 gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2521 tgcccgtgcc ctggcccacc ctcgtgacca ccttaggcta cggcgtggcc tgcttcgccc
     2581 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2641 tccaggagcg caccatctct ttcaaggacg acggtaccta caagacccgc gccgaggtga
     2701 agttcgaggg cgacaccctg gtgaaccgca tcgtgctgaa gggcatcgac ttcaaggagg
     2761 acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaag gtctatatca
     2821 cggccgacaa gcagaagaac ggcatcaagg ctaacttcaa gacccgccac aacgttgagg
     2881 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2941 tgctgctgcc cgacaaccac tacctgagcc atcagtccaa actgagcaaa gaccccaacg
     3001 agaagcgcga tcacatggtc ctgaaggaga gggtgaccgc cgccgggatt acacatgaca
     3061 tggacgagct gtacaagtaa caacgttgca cctgtgcctt ctagttgcca gccatctgtt
     3121 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3181 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3241 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3301 gcggtgggct ctatgg
//
