LOCUS       sequence-204516-        7255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRT integration of beta-globin reporter with premature termination 
            codon (PTC39); enables detection of 5'-3' decay intermediates 
            (xrFrag) and allows detection via northern blot (4H binding sites).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7255)
  AUTHORS   Boehm V, Gerbracht JV, Marx MC, Gehring NH
  TITLE     Interrogating the degradation pathways of unstable mRNAs with 
            XRN1-resistant sequences.
  JOURNAL   Nat Commun. 2016 Dec 5;7:13691. doi: 10.1038/ncomms13691.
  PUBMED    27917860
REFERENCE   2  (bases 1 to 7255)
  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: "Nat 
            Commun."; date: "2016-12-5"; volume: "7:13691. doi"; pages: " 
            10.1038/ncomms13691"
COMMENT     Sequence Label: pcDNA5/FRT/TO-globin_del5UTR_PTC39-xrRNA-4H
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    41..265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(398..417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    549..596
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             604..1626
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             604..1626
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    1756..1877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1793..1812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1847..1866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1926..1942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1926..1942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1939..1961)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1950..1966
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1974..1980,1981..1998,1999..2004))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2019..2040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2157..2174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2328..2916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2408..2427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3087..3878,3879..3947))
                     /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     3710..3729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3948..4052)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4127..4146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4318..4697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4698..4901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4852..4872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4903..4921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4924..4942
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4952..4976
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ggccgcagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
       61 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
      121 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
      181 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
      241 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
      301 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      541 cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg
      601 gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
      661 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
      721 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
      781 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
      841 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
      901 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
      961 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     1021 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     1081 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     1141 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     1201 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     1261 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     1321 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     1381 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     1441 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     1501 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     1561 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     1621 gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     1681 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1741 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1801 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1861 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     1921 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1981 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2041 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2101 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2161 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3001 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3061 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3121 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3181 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3241 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3301 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3361 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3421 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3481 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3541 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3601 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3661 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3721 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3781 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3841 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3901 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3961 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4021 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4081 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     4141 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     4201 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     4261 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     4321 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     4381 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4441 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4501 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4561 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4621 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4681 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4741 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4801 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4861 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag
     4921 atctccctat cagtgataga gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct
     4981 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     5041 ggactctagc gtttaaactt aagcttgcta gcaccatggt gcacctgact cctgaggaga
     5101 agtctgccgt tactgccctg tggggcaagg tgaacgtgga tgaagttggt ggtgaggccc
     5161 tgggcaggtt ggtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggcat
     5221 gtggagacag agaagactct tgggtttctg ataggcactg actctctctg cctattggtc
     5281 tattttccca cccttaggct gctggtggtc tacccttgga cctagaggtt ctttgagtcc
     5341 tttggggatc tgtccactcc tgatgctgtt atgggcaacc ctaaggtgaa ggctcatggc
     5401 aagaaagtgc tcggtgcctt tagtgatggc ctggctcacc tggacaacct caagggcacc
     5461 tttgccacac tgagtgagct gcactgtgac aagctgcacg tggatcctga gaacttcagg
     5521 gtgagtctat gggacccttg atgttttctt tccccttctt ttctatggtt aagttcatgt
     5581 cataggaagg ggagaagtaa cagggtacag tttagaatgg gaaacagacg aatgattgca
     5641 tcagtgtgga agtctcagga tcgttttagt ttcttttatt tgctgttcat aacaattgtt
     5701 ttcttttgtt taattcttgc tttctttttt tttcttctcc gcaattttta ctattatact
     5761 taatgcctta acattgtgta taacaaaagg aaatatctct gagatacatt aagtaactta
     5821 aaaaaaaact ttacacagtc tgcctagtac attactattt ggaatatatg tgtgcttatt
     5881 tgcatattca taatctccct actttatttt cttttatttt taattgatac ataatcatta
     5941 tacatattta tgggttaaag tgtaatgttt taatatgtgt acacatattg accaaatcag
     6001 ggtaattttg catttgtaat tttaaaaaat gctttcttct tttaatatac ttttttgttt
     6061 atcttatttc taatactttc cctaatctct ttctttcagg gcaataatga tacaatgtat
     6121 catgcctctt tgcaccattc taaagaataa cagtgataat ttctgggtta aggcaatagc
     6181 aatatttctg catataaata tttctgcata taaattgtaa ctgatgtaag aggtttcata
     6241 ttgctaatag cagctacaat ccagctacca ttctgctttt attttatggt tgggataagg
     6301 ctggattatt ctgagtccaa gctaggccct tttgctaatc atgttcatac ctcttatctt
     6361 cctcccacag ctcctgggca acgtgctggt ctgtgtgctg gcccatcact ttggcaaaga
     6421 attcacccca ccagtgcagg ctgcctatca gaaagtggtg gctggtgtgg ctaatgccct
     6481 ggcccacaag tatcactaag ctcgctttct tgctgtccaa tttctctcga ggtgagcaag
     6541 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtgg acttgatagt caggccagcc
     6601 ggttaggctg ccaccgaagg ttggtagacg gtgctgcctg cgaccaaccc caggaggact
     6661 gggttaccaa agctgattct ccacggttgg aaagcctccc agaaccgtct cggaagagga
     6721 gtccctgcca acaatggaga tgaagcccgt gtcagatcgc gaaagcgcca cttcgccgag
     6781 gagtgcaatc tgtgaggccc caggaggact gggtaaacag tcgagcaccc caccagtgca
     6841 ggctgcctat cagaaagtgg tggctggtgt ggctaatgcc ctggcccaca agtatcacta
     6901 agctcgcttt cttgctgtcc aatttgtcga gcaccccacc agtgcaggct gcctatcaga
     6961 aagtggtggc tggtgtggct aatgccctgg cccacaagta tcactaagct cgctttcttg
     7021 ctgtccaatt tgtcgagcac cccaccagtg caggctgcct atcagaaagt ggtggctggt
     7081 gtggctaatg ccctggccca caagtatcac taagctcgct ttcttgctgt ccaatttgtc
     7141 gagcacccca ccagtgcagg ctgcctatca gaaagtggtg gctggtgtgg ctaatgccct
     7201 ggcccacaag tatcactaag ctcgctttct tgctgtccaa tttgtcgacc cgggc
//
