LOCUS       (90-bp-TET-DIAL-        3736 bp DNA     circular SYN 18-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3736)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 18, 2025 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             118..927
                     /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    1001..1620
                     /label=pBR322_Origin
     rep_origin      1017..1605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      1770..1856
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1948..1975
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    2089..2107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2125..2143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2161..2179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2197..2215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2233..2251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2269..2287
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2305..2323
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2480..2513
                     /label=VloxP
     misc_feature    2605..2638
                     /label=VloxP
     protein_bind    complement(2659..2692)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2716..2740
                     /label=YB_TATA minimal promoter
     regulatory      2775..2784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2781..3500
                     /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    3512..3736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgctgactgc aggtgcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
       61 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
      121 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
      181 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
      241 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
      301 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
      361 atcaagcatt ttatccgtac tcctggtgat gcatggttac tcaccactgc gatccccgga
      421 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
      481 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
      541 gatcgcgtat ttcgtctagc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
      601 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
      661 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
      721 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
      781 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
      841 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
      901 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acattattca
      961 gattgggccc cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1681 tcccctgatt ctgtggataa ccgtattacc gctagcatgg atctcgggga cgtctaacta
     1741 ctaagcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
     1801 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1861 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1921 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
     1981 ttctacaaac tcttcctgtt agttagttac ttaagctcgg gcccacctgc agtctactgg
     2041 gaaccaattc agtcgactgg atccggtact actattcctc gagtttactc cctatcagtg
     2101 atagagaacg tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac
     2161 tccctatcag tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta
     2221 tgaccagttt actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg
     2281 atagagaacg tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc
     2341 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     2401 caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaac
     2461 tcgagtcgct cggtacgatt caatttctga gaactgtcat tctcggaaat tgagccaact
     2521 ttgtacaaaa aagcaggctc caccatggga accaattcag tcgactggat ccggtacctc
     2581 ctacagctcc tgggcaacgt gctgtcaatt tctgagaact gtcattctcg gaaattgaat
     2641 taattcgagc gaacgcgtat aacttcgtat agcatacatt atacgaagtt atcggatttg
     2701 atctgatcaa gcaactctag agggtatata atgggggcca ctagtctact accagagctc
     2761 atcgctagca atgagccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2821 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg
     2881 gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2941 tgcccgtgcc ctggcccacc ctcgtgacca ccttaggcta cggcgtggcc tgcttcgccc
     3001 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3061 tccaggagcg caccatctct ttcaaggacg acggtaccta caagacccgc gccgaggtga
     3121 agttcgaggg cgacaccctg gtgaaccgca tcgtgctgaa gggcatcgac ttcaaggagg
     3181 acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaag gtctatatca
     3241 cggccgacaa gcagaagaac ggcatcaagg ctaacttcaa gacccgccac aacgttgagg
     3301 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3361 tgctgctgcc cgacaaccac tacctgagcc atcagtccaa actgagcaaa gaccccaacg
     3421 agaagcgcga tcacatggtc ctgaaggaga gggtgaccgc cgccgggatt acacatgaca
     3481 tggacgagct gtacaagtaa caacgttgca cctgtgcctt ctagttgcca gccatctgtt
     3541 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3601 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3661 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3721 gcggtgggct ctatgg
//
