LOCUS       sequence-208352-        7043 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 7043)
  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 7043)
  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..7043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2761..2778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2767..2991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3124..3143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3275..3322
                     /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             3330..4352
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3330..4352
                     /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    4482..4603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4519..4538)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4573..4592
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4652..4668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4652..4668)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4665..4687)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4676..4692
                     /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(4700..4706,4707..4724,4725..4730))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4745..4766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4883..4900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5134..5153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5813..6604,6605..6673))
                     /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     6436..6455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6674..6778)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6853..6872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg ctagcaccat ggtgcacctg actcctgagg
      781 agaagtctgc cgttactgcc ctgtggggca aggtgaacgt ggatgaagtt ggtggtgagg
      841 ccctgggcag gttggtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
      901 catgtggaga cagagaagac tcttgggttt ctgataggca ctgactctct ctgcctattg
      961 gtctattttc ccacccttag gctgctggtg gtctaccctt ggacctagag gttctttgag
     1021 tcctttgggg atctgtccac tcctgatgct gttatgggca accctaaggt gaaggctcat
     1081 ggcaagaaag tgctcggtgc ctttagtgat ggcctggctc acctggacaa cctcaagggc
     1141 acctttgcca cactgagtga gctgcactgt gacaagctgc acgtggatcc tgagaacttc
     1201 agggtgagtc tatgggaccc ttgatgtttt ctttcccctt cttttctatg gttaagttca
     1261 tgtcatagga aggggataag taacagggta cagtttagaa tgggaaacag acgaatgatt
     1321 gcatcagtgt ggaagtctca ggatcgtttt agtttctttt atttgctgtt cataacaatt
     1381 gttttctttt gtttaattct tgctttcttt ttttttcttc tccgcaattt ttactattat
     1441 acttaatgcc ttaacattgt gtataacaaa aggaaatatc tctgagatac attaagtaac
     1501 ttaaaaaaaa actttacaca gtctgcctag tacattacta tttggaatat atgtgtgctt
     1561 atttgcatat tcataatctc cctactttat tttcttttat ttttaattga tacataatca
     1621 ttatacatat ttatgggtta aagtgtaatg ttttaatatg tgtacacata ttgaccaaat
     1681 cagggtaatt ttgcatttgt aattttaaaa aatgctttct tcttttaata tacttttttg
     1741 tttatcttat ttctaatact ttccctaatc tctttctttc agggcaataa tgatacaatg
     1801 tatcatgcct ctttgcacca ttctaaagaa taacagtgat aatttctggg ttaaggcaat
     1861 agcaatattt ctgcatataa atatttctgc atataaattg taactgatgt aagaggtttc
     1921 atattgctaa tagcagctac aatccagcta ccattctgct tttattttat ggttgggata
     1981 aggctggatt attctgagtc caagctaggc ccttttgcta atcatgttca tacctcttat
     2041 cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg ctggcccatc actttggcaa
     2101 agaattcacc ccaccagtgc aggctgccta tcagaaagtg gtggctggtg tggctaatgc
     2161 cctggcccac aagtatcact aagctcgctt tcttgctgtc caatttctct cgaggtgagc
     2221 aagggcgagg agctgttcac cggggtggtg cccatcctgg tggacttgat agtcaggcca
     2281 gccggttagg ctgccaccga aggttggtag acggtgctgc ctgcgaccaa ccccaggagg
     2341 actgggttac caaagctgat tctccacggt tggaaagcct cccagaaccg tctcggaaga
     2401 ggagtccctg ccaacaatgg agatgaagcc cgtgtcagat cgcgaaagcg ccacttcgcc
     2461 gaggagtgca atctgtgagg ccccaggagg actgggtaaa cagtcgagca ccccaccagt
     2521 gcaggctgcc tatcagaaag tggtggctgg tgtggctaat gccctggccc acaagtatca
     2581 ctaagctcgc tttcttgctg tccaatttgt cgagcacccc accagtgcag gctgcctatc
     2641 agaaagtggt ggctggtgtg gctaatgccc tggcccacaa gtatcactaa gctcgctttc
     2701 ttgctgtcca atttgtcgac ccgggcggcc gcagagggcc cgtttaaacc cgctgatcag
     2761 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2821 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2881 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2941 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3001 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3241 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     3301 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     3361 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     3421 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     3481 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     3541 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     3601 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     3661 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     3721 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     3781 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     3841 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3901 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3961 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     4021 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     4081 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     4141 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     4201 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     4261 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     4321 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     4381 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4441 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4501 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4561 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4621 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6781 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6901 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6961 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     7021 tgtacgggcc agatatacgc gtt
//
