LOCUS       sequence-169938-        6054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  beta Globin NMD construct with and without PTC under Bi-directional 
            TET on promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6054)
  AUTHORS   Trcek T, Sato H, Singer RH, Maquat LE
  TITLE     Temporal and spatial characterization of nonsense-mediated mRNA 
            decay.
  JOURNAL   Genes Dev. 2013 Mar 1;27(5):541-51. doi: 10.1101/gad.209635.112. 
            Epub 2013 Feb 21.
  PUBMED    23431032
REFERENCE   2  (bases 1 to 6054)
  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"; doi: 
            "10.1101/gad.209635.112"; journalName: "Genes Dev"; date: 
            "2013-03-1- 1"; volume: "27"; issue: "5"; pages: "541-51"
COMMENT     Sequence Label: pTRE-Tight-BI-Gl NORM-LacZA-TER-LacZB
FEATURES             Location/Qualifiers
     source          1..6054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     polyA_signal    2123..2204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2215..2232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2386..2974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2466..2485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3145..3936,3937..4005))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3768..3787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4006..4110)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    4143..4224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        join(5985..6054,1..318)
                     /label=bidirectional TRE promoter
                     /note="Tet-responsive bidirectional promoter PTight-BI, 
                     consisting of seven tet operator sequences flanked on each 
                     side by the minimal CMV promoter"
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 gagatttcga gctcggtacc cggggatcct ctagtcaggg
      361 ggatggtgca cctgactcct gaggagaagt ctgccgttac tgccctgtgg ggcaaggtga
      421 acgtggatga agttggtggt gaggccctgg gcaggttggt atcaaggtta caagacaggt
      481 ttaaggagac caatagaaac tgggcatgtg gagacagaga agactcttgg gtttctgata
      541 ggcactgact ctctctgcct attggtctat tttcccaccc ttaggctgct ggtggtctac
      601 ccttggaccc agaggttctt tgagtccttt ggggatctgt ccactcctga tgctgttatg
      661 ggcaacccta aggtgaaggc tcatggcaag aaagtgctcg gtgcctttag tgatggcctg
      721 gctcacctgg acaacctcaa gggcaccttt gccacactga gtgagctgca ctgtgacaag
      781 ctgcacgtgg atcctgagaa cttcagggtg agtctgatgg gcacctcctg ggtttccttc
      841 ccctggctat tctgctcaac cttcctatca gaaaaaaagg ggaagcgatt ctagggagca
      901 gtctccatga ctgtgtgtgg agtgttgaca agagttcgga tattttattc tctactcaga
      961 attgctgctc cccctcactc tgttctgtgt tgtcatttcc tctttctttg gtaagctttt
     1021 aatttccagt tgcattttac taaattaatt aagctggtta tttacttccc atcctgatat
     1081 cagcttcccc tcctcctttc ctcccagtcc ttctctctct cctctctctt tctctaatcc
     1141 tttcctttcc ctcagttcat ttcttcttct ttgatctacg tttgtttgtc tttttaaata
     1201 ttgccttgta acttgctcag aggacaagga agatatgtcc ctgtttcttc tcatagctct
     1261 caagaatagt agcataattg gcttttatgc cagggtgaca ggggaagaat atattttaca
     1321 tataaattct gtttgacata ggattcttat aataatttgt cagtagttta aggttgcaaa
     1381 caaatgtctt tgtaaataag cctgcagtat ctggtatttt tgctctacag ttatgttgat
     1441 ggttcttcca tattcccaca gctcctgggc aatatgatcg tgattgtgct gggccaccac
     1501 ctgggcaagg atttcacccc cgctgcacag gctgccttcc agaaggtggt ggctggagtg
     1561 gccactgccc tggctcacaa gtaccactaa acccggtacc aggccagacg cgaattattt
     1621 ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc ggttacggcc
     1681 aggacagtcg tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc ggagaaaacc
     1741 gcctcgcggt gatggtgctg cgctggagtg acggcagtta tctggaagat caggatatgt
     1801 ggcggatgag cggcattttc cgtgacgtct cgttgctgca taaaccgact acacaaatca
     1861 gcgatttcca tgttgccact cgctttaatg atgatttcag ccgcgctgta ctggaggctg
     1921 aagttcagat ggtaccgtga gggttcaggt ctgaccttgg ggctgacgcg tgctagcgcg
     1981 gccgcatcga taagcttgtc gacgatatct ccagaggatc ataatcagcc ataccacatt
     2041 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2101 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2161 caatagcatc acaaatttca caaataaagc atttttttca ctgccccgag cttcctcgct
     2221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2821 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3001 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     3781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4141 cggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4201 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4261 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     4321 gattatgatc ctctagactg cagcctcagg agatctcccc aaggtcagac tgaacccaca
     4381 cggtaccgct ggatgcggcg tgcggtcggc aaagaccaga ccgttcatac agaactggcg
     4441 atcgttcggc gtatcgccaa aatcaccgcc gtaagccgac cacgggttgc cgttttcatc
     4501 atatttaatc agcgactgat ccacccagtc ccagacgaag tcgccctgta aacggggata
     4561 ctgacgaaac gcctgccagt atttagcgaa accgccaaga ctgttaccca tcgcgtgggc
     4621 gtattcgcaa aggatcagcg ggcgcgtctc tccaggtagc gaaagccatt ttttgatgga
     4681 ccatttcggc acagccggga agggctggtc ttcatccggt accgggttta gtggtacttg
     4741 tgagccaggg cagtggccac tccagccacc accttctgga aggcagcctg tgcagcgggg
     4801 gtgaaatcct tgcccaggtg gtggcccagc acaatcacga tcatattgcc caggagctgt
     4861 gggaatatgg aagaaccatc aacataactg tagagcaaaa ataccagata ctgcaggctt
     4921 atttacaaag acatttgttt gcaaccttaa actactgaca aattattata agaatcctat
     4981 gtcaaacaga atttatatgt aaaatatatt cttcccctgt caccctggca taaaagccaa
     5041 ttatgctact attcttgaga gctatgagaa gaaacaggga catatcttcc ttgtcctctg
     5101 agcaagttac aaggcaatat ttaaaaagac aaacaaacgt agatcaaaga agaagaaatg
     5161 aactgaggga aaggaaagga ttagagaaag agagaggaga gagagaagga ctgggaggaa
     5221 aggaggaggg gaagctgata tcaggatggg aagtaaataa ccagcttaat taatttagta
     5281 aaatgcaact ggaaattaaa agcttaccaa agaaagagga aatgacaaca cagaacagag
     5341 tgagggggag cagcaattct gagtagagaa taaaatatcc gaactcttgt caacactcca
     5401 cacacagtca tggagactgc tccctagaat cgcttcccct ttttttctga taggaaggtt
     5461 gagcagaata gccaggggaa ggaaacccag gaggtgccca tcagactcac cctgaagttc
     5521 tcaggatcca cgtgcagctt gtcacagtgc agctcactca gtgtggcaaa ggtgcccttg
     5581 aggttgtcca ggtgagccag gccatcacta aaggcaccga gcactttctt gccatgagcc
     5641 ttcaccttag ggttgcccat aacagcatca ggagtggaca gatccccaaa ggactcaaag
     5701 aacctctagg tccaagggta gaccaccagc agcctaaggg tgggaaaata gaccaatagg
     5761 cagagagagt cagtgcctat cagaaaccca agagtcttct ctgtctccac atgcccagtt
     5821 tctattggtc tccttaaacc tgtcttgtaa ccttgatacc aacctgccca gggcctcacc
     5881 accaacttca tccacgttca ccttgcccca cagggcagta acggcagact tctcctcagg
     5941 agtcaggtgc accatggcat atgccctgac cggtgaattc tccaggcgat ctgacggttc
     6001 actaaacgag ctctgcttat ataggcctcc caccgtacac gccacctcga cata
//
