LOCUS       sequence-8174-ex        3087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3087)
  AUTHORS   Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP, Burge CB
  TITLE     Prediction of mammalian microRNA targets.
  JOURNAL   Cell. 2003 Dec 26. 115(7):787-98.
  PUBMED    14697198
REFERENCE   2  (bases 1 to 3087)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2003 
            Dec 26. 115(7):787-98."
COMMENT     Sequence Label: pTRE-TIGHT miR-1 (aka:pTRE-m1d)
FEATURES             Location/Qualifiers
     source          1..3087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     291..315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    1001..1082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1038..1057)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1093..1110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1264..1852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1344..1363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2023..2814,2815..2883))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2646..2665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2884..2988)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3056..3074
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
       61 tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
      121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
      181 tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
      241 gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta
      301 gtgaaccgtc agatcgcctg gagaattcga gctcggtacc cggggatccg agagatggat
      361 tcagggatgg aggggcgccg aggtcagaag ggggcaggga gtcctgggcc cctgggggtg
      421 gctggacacc aggcagcagt ggcaggaagg ctgtcctgct cacacagaga gggctccggc
      481 agtagactcc agggaagaag ttacactgcc tctgagctgc cttctctaca tcgcagtggg
      541 gtcagcttct accggggcgg cgtcccgggg tcttggaact gcatgcagac tgcctgcttg
      601 ggaaacatac ttctttatat gcccatatgg acctgctaag ctatggaatg taaagaagta
      661 tgtatctcag gccgggacct ctctcgccgc actgaggggc actccacacc acgggggccg
      721 ccgcggccga cagccagcca ggcggctgtg gtgggggtga ggctggggcg gcaggctgcc
      781 tgggcgttgg tggacggtcg tgtccccgcc agcattgttc tgtcactttg cctgtgtcag
      841 caggtggaag tgctcaccag acaatcgata agcttgtcga cgatatctct agaggatcat
      901 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
      961 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1021 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1081 gcctcgagct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1441 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1681 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     1741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3001 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3061 gcgtatcacg aggccctttc gtcttca
//
