LOCUS       sequence-208932-        6060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of Pumilio1, fused to iRFP670 in mammalian 
            cells, under a hUBC promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6060)
  AUTHORS   Maass PG, Barutcu AR, Shechner DM, Weiner CL, Mele M, Rinn JL
  TITLE     Spatiotemporal allele organization by allele-specific CRISPR 
            live-cell imaging (SNP-CLING).
  JOURNAL   Nat Struct Mol Biol. 2018 Jan 8. pii: 10.1038/s41594-017-0015-3. 
            doi: 10.1038/s41594-017-0015-3.
  PUBMED    29343869
REFERENCE   2  (bases 1 to 6060)
  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 Struct 
            Mol Biol. 2018 Jan 8. pii: 10.1038/s41594-017-0015-3. doi: 
            10.1038/s41594-017-0015-3."
COMMENT     Sequence Label: pUBC_hsPUM1-iRFP670
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1056..1075
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1269..1291
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1283..1300
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1284..1300
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1329..2540
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2377..2396
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      2558..2567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2570..2590
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2597..2623
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3788..3797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3794..4729
                     /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"
     primer_bind     complement(4770..4789)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4824..4843
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4900..4916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4900..4916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4913..4935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4924..4940
                     /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(4948..4954,4955..4972,4973..4978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4993..5014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5131..5148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5302..5890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5382..5401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1021 attaacctat aaaaataggc gtatcacgag ctatgcggca tcagagcaga ttgtactgag
     1081 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     1141 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     1201 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     1261 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacacg
     1321 tatgtcgagg taggcgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc
     1381 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct
     1441 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     1501 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     1561 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     1621 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     1681 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     1741 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     1801 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     1861 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     1921 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     1981 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg
     2041 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg
     2101 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc
     2161 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg
     2221 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc
     2281 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc
     2341 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc
     2401 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat
     2461 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt
     2521 ttttggcttt tttgttagac ctggaggtac cgccaccgcc accatggctc caaagaagaa
     2581 gcgtaaggta ggcagctacc catacgatgt tccagattac gctggcagcg gtcggtcccg
     2641 gctcttggag gactttcgaa acaatcgata tcctaacttg cagcttcgcg agatcgctgg
     2701 tcatattatg gaattctctc aggatcaaca cgggtctaga ttcattcagc tcaagctgga
     2761 gagagccaca cctgccgaac ggcagctcgt gtttaatgag atcctccagg ctgcctatca
     2821 actgatggtg gacgtgttcg gcaattacgt catccagaag ttctttgaat ttggtagtct
     2881 cgagcagaag ctggcccttg cagagagaat ccgcggccat gtactgtccc ttgccctgca
     2941 gatgtacggg tgccgcgtaa tccagaaggc actggaattt ataccgtccg accagcagaa
     3001 tgagatggtg cgggaactcg atggtcatgt gttgaaatgc gtaaaagatc agaacggtaa
     3061 ccacgtggtt cagaaatgta tcgagtgtgt ccagccacag agcttgcaat tcatcataga
     3121 tgcctttaag ggccaggttt tcgctctgag cacgcacccc tacggatgcc gggtcattca
     3181 gcggattctt gaacattgtc tgccagacca gactctgcct atcctggagg agctccacca
     3241 acacaccgaa cagttggtcc aggatcaata tggtaattac gttattcagc atgtgctgga
     3301 acatgggcgg cctgaggaca agtccaaaat tgtagctgag attcgaggca atgtgctcgt
     3361 tctttcccag cacaagttcg cttctaacgt cgtggagaaa tgtgtgactc acgccagccg
     3421 gactgagcgc gcggtgctca ttgatgaagt gtgtaccatg aacgatgggc cacatagcgc
     3481 actttacaca atgatgaaag accagtacgc aaactacgtg gttcagaaga tgatagacgt
     3541 cgctgaaccg gggcagcgca aaattgtcat gcacaagatc cgcccacaca tagctactct
     3601 tcggaaatac acctatggca agcacatcct cgcaaagctg gagaagtatt acatgaaaaa
     3661 cggggtggat ctgggaggat ccaagggctc gggctcgacc tcgggctccg gaatgtcgcg
     3721 gaggcggcac agctacgaga acgatggtgg acaacctcac aaaaggagga agacgtctcc
     3781 accggtcgcc accatggcgc gtaaggtcga tctcacctcc tgcgatcgcg agccgatcca
     3841 catccccggc agcattcagc cgtgcggctg cctgctagcc tgcgacgcgc aggcggtgcg
     3901 gatcacgcgc attacggaaa atgccggcgc gttctttgga cgcgaaactc cgcgggtcgg
     3961 tgagctactc gccgattact tcggcgagac cgaagcccat gcgctgcgca acgcactggc
     4021 gcagtcctcc gatccaaagc gaccggcgct gatcttcggt tggcgcgacg gcctgaccgg
     4081 ccgcaccttc gacatctcac tgcatcgcca tgacggtaca tcgatcatcg agttcgagcc
     4141 tgcggcggcc gaacaggccg acaatccgct gcggctgacg cggcagatca tcgcgcgcac
     4201 caaagaactg aagtcgctcg aagagatggc cgcacgggtg ccgcgctatc tgcaggcgat
     4261 gctcggctat caccgcgtga tgttgtaccg cttcgcggac gacggctccg ggatggtgat
     4321 cggcgaggcg aagcgcagcg acctcgagag ctttctcggt cagcactttc cggcgtcgct
     4381 ggtcccgcag caggcgcggc tactgtactt gaagaacgcg atccgcgtgg tctcggattc
     4441 gcgcggcatc agcagccgga tcgtgcccga gcacgacgcc tccggcgccg cgctcgatct
     4501 gtcgttcgcg cacctgcgca gcatctcgcc ctgccatctc gaatttctgc ggaacatggg
     4561 cgtcagcgcc tcgatgtcgc tgtcgatcat cattgacggc acgctatggg gattgatcat
     4621 ctgtcatcat tacgagccgc gtgccgtgcc gatggcgcag cgcgtcgcgg ccgaaatgtt
     4681 cgccgacttc ttatcgctgc acttcaccgc cgcccaccac caacgctaag tcgacttgtt
     4741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttattcgac
     4861 tgtttaaacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
//
