LOCUS       (203-bp-DIAL-min        3501 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 3501)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3501
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        1..39
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    40..64
                     /label=ZF43/ZF37_hybridpromoter
     regulatory      70..79
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(76..78,79..81,82..795)
                     /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    807..1031
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1054..1077
                     /label=pENTR2B.Seq.REV
     CDS             1153..1962
                     /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    2036..2655
                     /label=pBR322_Origin
     rep_origin      2052..2640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      2805..2891
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2983..3010
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    3087..3189
                     /label=ZF43/ZF37_hybridpromoter
     protein_bind    3099..3107
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3103..3111
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3114..3122
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3118..3126
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3129..3137
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3133..3141
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3144..3152
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3148..3156
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3159..3167
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3163..3171
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    3174..3182
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    3178..3186
                     /label=ZF37 Binding Site
                     /bound_moiety="ZF37"
     protein_bind    complement(3209..3242)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3446..3479)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ggtaggcgtg tacggtggga ggcctatata agcagagctt ctactaccag agctcatcgc
       61 tagcaatgag ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      121 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtccgcgg cgagggcgag
      181 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      241 gtgccctggc ccaccctcgt gaccacctta ggctacggcg tggcctgctt cgcccgctac
      301 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      361 gagcgcacca tctctttcaa ggacgacggt acctacaaga cccgcgccga ggtgaagttc
      421 gagggcgaca ccctggtgaa ccgcatcgtg ctgaagggca tcgacttcaa ggaggacggc
      481 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaaggtcta tatcacggcc
      541 gacaagcaga agaacggcat caaggctaac ttcaagaccc gccacaacgt tgaggacggc
      601 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      661 ctgcccgaca accactacct gagccatcag tccaaactga gcaaagaccc caacgagaag
      721 cgcgatcaca tggtcctgaa ggagagggtg accgccgccg ggattacaca tgacatggac
      781 gagctgtaca agtaacaacg ttgcacctgt gccttctagt tgccagccat ctgttgtttg
      841 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
      901 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
      961 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1021 gggctctatg gcgctcgctg actgcaggtg cccgtgtctc aaaatctctg atgttacatt
     1081 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1141 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     1201 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1261 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1321 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1381 atgcctcttc cgaccatcaa gcattttatc cgtactcctg gtgatgcatg gttactcacc
     1441 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     1501 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     1561 tgtcctttta acagcgatcg cgtatttcgt ctagctcagg cgcaatcacg aatgaataac
     1621 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     1681 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     1741 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     1801 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     1861 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     1921 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     1981 ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt agaaaagatc
     2041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag catggatctc
     2761 ggggacgtct aactactaag cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     2821 ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     2881 gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc
     2941 gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg
     3001 atggcctttt tgcgtttcta caaactcttc ctgttagtta gttacttaag ctcgggccca
     3061 cctgcagtct acttactcgg atcgggagat ctgaattgga gtgaggacgt gtagagtgag
     3121 gacgtgcgga gtgaggacgt gtagagtgag gacgtgcgga gtgaggacgt gtagagtgag
     3181 gacgtgccct cctctggatc aaatccgaat aacttcgtat agcatacatt atacgaagtt
     3241 atctcgagtc gctcggtacg atttaaattg aattctgcag tcgacggtac cgcgggcccg
     3301 ggactttgta caaaaaagca ggctccacca tgggaaccaa ttcagtcgac tggatccggt
     3361 acctcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     3421 aattgattaa ttcgagcgaa cgcgtataac ttcgtatagc atacattata cgaagttatc
     3481 ggatttgatc tgatcaagca a
//
