LOCUS       sequence-161547-        1012 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses CRE and iRFP670 under two separate PGK promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1012)
  AUTHORS   Daneshvar K, Pondick JV, Kim BM, Zhou C, York SR, Macklin JA, 
            Abualteen A, Tan B, Sigova AA, Marcho C, Tremblay KD, Mager J, Choi 
            MY, Mullen AC
  TITLE     DIGIT Is a Conserved Long Noncoding RNA that Regulates GSC 
            Expression to Control Definitive Endoderm Differentiation of 
            Embryonic Stem Cells.
  JOURNAL   Cell Rep. 2016 Oct 4;17(2):353-365. doi: 
            10.1016/j.celrep.2016.09.017.
  PUBMED    27705785
REFERENCE   2  (bases 1 to 1012)
  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: 
            "10.1016/j.celrep.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-4- 4"; volume: "17"; issue: "2"; pages: "353-365"
COMMENT     Sequence Label: pCRE-iRFP670
FEATURES             Location/Qualifiers
     source          1..1012
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(109..129)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(154..653)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(202..221)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     595..618
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(678..694)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(679..695)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(723..741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(762..778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(762..778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(775..797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    786..802
                     /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(810..816,817..834,835..840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    855..876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(993..1010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaacgcctgg cgatccctga acatgtccat caggttcttg cgaacctcat cactcgttgc
       61 atcgaccggt aatgcaggca aattttggtg tacggtcagt aaattggaca ccttcctctt
      121 cttcttgggc atggtcgacc tcgagctgca ggtcgaaagg cccggagatg aggaagagga
      181 gaacagcgcg gcagacgtgc gcttttgaag cgtgcagaat gccgggcctc cggaggacct
      241 tcgggcgccc gccccgcccc tgagcccgcc cctgagcccg cccccggacc caccccttcc
      301 cagcctctga gcccagaaag cgaaggagca aagctgctat tggccgctgc cccaaaggcc
      361 tacccgcttc cattgctcag cggtgctgtc catctgcacg agactagtga gacgtgctac
      421 ttccatttgt cacgtcctgc acgacgcgag ctgcggggcg ggggggaact tcctgactag
      481 gggaggagta gaaggtggcg cgaaggggcc accaaagaac ggagccggtt ggcgcctacc
      541 ggtggatgtg gaatgtgtgc gaggccagag gccacttgtg tagcgccaag tgcccagcgg
      601 ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc gcctccccta cccggtagaa
      661 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac cagcttttgt
      721 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
      781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      841 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gc
//
