LOCUS       sequence-158775-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of coding/noncoding genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  TITLE     pTREtight reporter plasmids--Roderic Guigo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4767)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTre-tight-SV40-luciferase (496)
FEATURES             Location/Qualifiers
     source          1..4767
                     /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    519..600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(611..628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(782..1370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(862..881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1541..2332,2333..2401))
                     /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     2164..2183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2402..2506)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2517..2713
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2564..2699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2749..2758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2755..4407
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    4451..4572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4462..4481)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4736..4754
                     /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 cggggatcct ctagtcagct
      361 gacgcgtgct agcgcggccg catcgataag cttgtcgacg atatctctag aggatcataa
      421 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
      481 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
      541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
      601 ctcgagcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      661 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      721 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      781 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      961 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1201 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     1261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1381 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1441 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccctgca
     2521 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2581 gcccagttcc gcccattctc cgccccatcg ctgactaatt ttttttattt atgcagaggc
     2641 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2701 aggcttttgc aaaaagctta agcttggcaa tccggtactg ttggtaaagc caccatggaa
     2761 gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc
     2821 ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt
     2881 accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg
     2941 ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc
     3001 gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg
     3061 gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag
     3121 cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag
     3181 ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa
     3241 agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg
     3301 cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc
     3361 ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc
     3421 cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca
     3481 tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc
     3541 gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt
     3601 caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac
     3661 aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag
     3721 gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg
     3781 acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta
     3841 ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg
     3901 ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt
     3961 aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac
     4021 atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc
     4081 aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc
     4141 aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc
     4201 gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg
     4261 gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg
     4321 cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc
     4381 aagaagggcg gcaagatcgc cgtgtaataa ttctagagtc ggggcggccg gccgcttcga
     4441 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4501 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4561 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     4621 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     4681 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4741 gcgtatcacg aggccctttc gtcttca
//
