LOCUS       sequence-193616-        3835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  theophylline-agRNA-FANCF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3835)
  AUTHORS   Tang W, Hu JH, Liu DR
  TITLE     Aptazyme-embedded guide RNAs enable ligand-responsive genome editing
            and transcriptional activation.
  JOURNAL   Nat Commun. 2017 Jun 28;8:15939. doi: 10.1038/ncomms15939.
  PUBMED    28656978
REFERENCE   2  (bases 1 to 3835)
  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: "2017-06-28"; volume: "8:15939. doi"; pages: " 
            10.1038/ncomms15939"
COMMENT     Sequence Label: pWT055b
FEATURES             Location/Qualifiers
     source          1..3835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        448..523
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        587..686
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     637..657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    709..789
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      791..800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             797..1732
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    1854..1975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1891..1910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1945..1964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2086..2674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2166..2185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2845..3636,3637..3705))
                     /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     3468..3487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3706..3810)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gctgcagaag ggattccggt acatccagct
      361 gatgagtccc aaataggacg aaatacatac cagccgaaag gcccttggca ggtgtcctgg
      421 attccacgga atcccttctg cagcaccgtt ttagagctag aaatagcaag ttaaaataag
      481 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt aagcttgggc
      541 cgctcgaggt acctctctac atatgacatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      721 ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
      781 tccaccggtc gccaccatgg cgcgtaaggt cgatctcacc tcctgcgatc gcgagccgat
      841 ccacatcccc ggcagcattc agccgtgcgg ctgcctgcta gcctgcgacg cgcaggcggt
      901 gcggatcacg cgcattacgg aaaatgccgg cgcgttcttt ggacgcgaaa ctccgcgggt
      961 cggtgagcta ctcgccgatt acttcggcga gaccgaagcc catgcgctgc gcaacgcact
     1021 ggcgcagtcc tccgatccaa agcgaccggc gctgatcttc ggttggcgcg acggcctgac
     1081 cggccgcacc ttcgacatct cactgcatcg ccatgacggt acatcgatca tcgagttcga
     1141 gcctgcggcg gccgaacagg ccgacaatcc gctgcggctg acgcggcaga tcatcgcgcg
     1201 caccaaagaa ctgaagtcgc tcgaagagat ggccgcacgg gtgccgcgct atctgcaggc
     1261 gatgctcggc tatcaccgcg tgatgttgta ccgcttcgcg gacgacggct ccgggatggt
     1321 gatcggcgag gcgaagcgca gcgacctcga gagctttctc ggtcagcact ttccggcgtc
     1381 gctggtcccg cagcaggcgc ggctactgta cttgaagaac gcgatccgcg tggtctcgga
     1441 ttcgcgcggc atcagcagcc ggatcgtgcc cgagcacgac gcctccggcg ccgcgctcga
     1501 tctgtcgttc gcgcacctgc gcagcatctc gccctgccat ctcgaatttc tgcggaacat
     1561 gggcgtcagc gcctcgatgt cgctgtcgat catcattgac ggcacgctat ggggattgat
     1621 catctgtcat cattacgagc cgcgtgccgt gccgatggcg cagcgcgtcg cggccgaaat
     1681 gttcgccgac ttcttatcgc tgcacttcac cgccgcccac caccaacgct aagcggccgc
     1741 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1801 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     1981 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttatgt gagcaaaagg
     2041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacct
//
