LOCUS       sequence-228522-        2932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows in vitro SP6 Polymerase driven transcription of a beta globin
            northern blot probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2932)
  AUTHORS   Voigt F, Gerbracht JV, Boehm V, Horvathova I, Eglinger J, Chao JA, 
            Gehring NH
  TITLE     Detection and quantification of RNA decay intermediates using 
            XRN1-resistant reporter transcripts.
  JOURNAL   Nat Protoc. 2019 May;14(5):1603-1633. doi: 
            10.1038/s41596-019-0152-8. Epub 2019 Apr 24.
  PUBMED    31019309
REFERENCE   2  (bases 1 to 2932)
  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: 
            "10.1038/s41596-019-0152-8"; journalName: "Nat Protoc"; date: 
            "2019-05"; volume: "14"; issue: "5"; pages: "1603-1633"
COMMENT     Sequence Label: pGEM4Z globin NB Probe
FEATURES             Location/Qualifiers
     source          1..2932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(254..273)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(255..273)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(292..308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(292..308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(305..327)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    316..332
                     /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(340..346,347..364,365..370))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    385..406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(523..540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(694..1282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(774..793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1453..2244,2245..2313))
                     /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     2076..2095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2314..2418)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2486..2504
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2542..2564)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2664..2683
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2877..2899
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2891..2908
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2892..2908
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(2916..2932,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(2916..2932,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaatacgaat tcgagctcgg tacccgggca aaggaacctt taatagaaat tggacagcaa
       61 gaaagcgagc ttagtgatac ttgtgggcca gggcattagc cacaccagcc accactttct
      121 gataggcagc ctgcactggt ggggtgaatt ctttgccaaa gtgatgggcc agcacacaga
      181 ccagcacgtt gcccaggagc ctgaagttct cagggatcct ctagagtcga cctgcaggca
      241 tgcaagcttg tctccctata gtgagtcgta ttagagcttg gcgtaatcat ggtcatagct
      301 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      361 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      421 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      481 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
      541 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      601 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     2461 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     2521 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2581 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2641 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     2701 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     2761 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2821 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     2881 gtcacgacgt tgtaaaacga cggccagtga attggattta ggtgacacta ta
//
