LOCUS       sequence-233254-        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     66..88
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     80..97
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     81..97
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        105..123
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     105..122
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(375..394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(376..394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(413..429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(413..429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(426..448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    437..453
                     /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(461..467,468..485,486..491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    506..527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(644..661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(815..1403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(895..914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1574..2365,2366..2434))
                     /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     2197..2216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2435..2539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2607..2625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2663..2685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2785..2804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
       61 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttggatttag gtgacactat
      121 agaatacgaa ttcgagctcg gtacccgggc aaaggaacct ttaatagaaa ttggacagca
      181 agaaagcgag cttagtgata cttgtgggcc agggcattag ccacaccagc caccactttc
      241 tgataggcag cctgcactgg tggggtgaat tctttgccaa agtgatgggc cagcacacag
      301 accagcacgt tgcccaggag cctgaagttc tcagggatcc tctagagtcg acctgcaggc
      361 atgcaagctt gtctccctat agtgagtcgt attagagctt ggcgtaatca tggtcatagc
      421 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      481 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      541 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2581 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2641 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2701 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2761 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2821 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2881 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ct
//
