LOCUS       sequence-207515-        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(19..810,811..879))
                     /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     642..661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(880..984)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1074..1093
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1287..1309
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1301..1318
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1302..1318
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1347..2558
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2395..2414
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      2576..2585
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2588..2608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2615..2641
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3806..3815
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3812..4747
                     /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(4788..4807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4842..4861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4918..4934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4918..4934)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4931..4953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4942..4958
                     /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(4966..4972,4973..4990,4991..4996))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5011..5032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5149..5166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5320..5908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5400..5419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
       61 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
      481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
      661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      961 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     1021 aaccattatt atcatgacat taacctataa aaataggcgt atcacgagct atgcggcatc
     1081 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     1141 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     1201 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     1261 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     1321 attcgagctc ggtacacgta tgtcgaggta ggcgcgccgg gttttggcgc ctcccgcggg
     1381 cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga
     1441 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     1501 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     1561 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     1621 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     1681 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     1741 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     1801 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     1861 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     1921 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     1981 gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat
     2041 gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     2101 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     2161 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     2221 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     2281 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     2341 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     2401 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     2461 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt
     2521 ccgctaaatt ctggccgttt ttggcttttt tgttagacct ggaggtaccg ccaccgccac
     2581 catggctcca aagaagaagc gtaaggtagg cagctaccca tacgatgttc cagattacgc
     2641 tggcagcggt cggtcccggc tcttggagga ctttcgaaac aatcgatatc ctaacttgca
     2701 gcttcgcgag atcgctggtc atattatgga attctctcag gatcaacacg ggtctagatt
     2761 cattcagctc aagctggaga gagccacacc tgccgaacgg cagctcgtgt ttaatgagat
     2821 cctccaggct gcctatcaac tgatggtgga cgtgttcggc aattacgtca tccagaagtt
     2881 ctttgaattt ggtagtctcg agcagaagct ggcccttgca gagagaatcc gcggccatgt
     2941 actgtccctt gccctgcaga tgtacgggtg ccgcgtaatc cagaaggcac tggaatttat
     3001 accgtccgac cagcagaatg agatggtgcg ggaactcgat ggtcatgtgt tgaaatgcgt
     3061 aaaagatcag aacggtaacc acgtggttca gaaatgtatc gagtgtgtcc agccacagag
     3121 cttgcaattc atcatagatg cctttaaggg ccaggttttc gctctgagca cgcaccccta
     3181 cggatgccgg gtcattcagc ggattcttga acattgtctg ccagaccaga ctctgcctat
     3241 cctggaggag ctccaccaac acaccgaaca gttggtccag gatcaatatg gtaattacgt
     3301 tattcagcat gtgctggaac atgggcggcc tgaggacaag tccaaaattg tagctgagat
     3361 tcgaggcaat gtgctcgttc tttcccagca caagttcgct tctaacgtcg tggagaaatg
     3421 tgtgactcac gccagccgga ctgagcgcgc ggtgctcatt gatgaagtgt gtaccatgaa
     3481 cgatgggcca catagcgcac tttacacaat gatgaaagac cagtacgcaa actacgtggt
     3541 tcagaagatg atagacgtcg ctgaaccggg gcagcgcaaa attgtcatgc acaagatccg
     3601 cccacacata gctactcttc ggaaatacac ctatggcaag cacatcctcg caaagctgga
     3661 gaagtattac atgaaaaacg gggtggatct gggaggatcc aagggctcgg gctcgacctc
     3721 gggctccgga atgtcgcgga ggcggcacag ctacgagaac gatggtggac aacctcacaa
     3781 aaggaggaag acgtctccac cggtcgccac catggcgcgt aaggtcgatc tcacctcctg
     3841 cgatcgcgag ccgatccaca tccccggcag cattcagccg tgcggctgcc tgctagcctg
     3901 cgacgcgcag gcggtgcgga tcacgcgcat tacggaaaat gccggcgcgt tctttggacg
     3961 cgaaactccg cgggtcggtg agctactcgc cgattacttc ggcgagaccg aagcccatgc
     4021 gctgcgcaac gcactggcgc agtcctccga tccaaagcga ccggcgctga tcttcggttg
     4081 gcgcgacggc ctgaccggcc gcaccttcga catctcactg catcgccatg acggtacatc
     4141 gatcatcgag ttcgagcctg cggcggccga acaggccgac aatccgctgc ggctgacgcg
     4201 gcagatcatc gcgcgcacca aagaactgaa gtcgctcgaa gagatggccg cacgggtgcc
     4261 gcgctatctg caggcgatgc tcggctatca ccgcgtgatg ttgtaccgct tcgcggacga
     4321 cggctccggg atggtgatcg gcgaggcgaa gcgcagcgac ctcgagagct ttctcggtca
     4381 gcactttccg gcgtcgctgg tcccgcagca ggcgcggcta ctgtacttga agaacgcgat
     4441 ccgcgtggtc tcggattcgc gcggcatcag cagccggatc gtgcccgagc acgacgcctc
     4501 cggcgccgcg ctcgatctgt cgttcgcgca cctgcgcagc atctcgccct gccatctcga
     4561 atttctgcgg aacatgggcg tcagcgcctc gatgtcgctg tcgatcatca ttgacggcac
     4621 gctatgggga ttgatcatct gtcatcatta cgagccgcgt gccgtgccga tggcgcagcg
     4681 cgtcgcggcc gaaatgttcg ccgacttctt atcgctgcac ttcaccgccg cccaccacca
     4741 acgctaagtc gacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4801 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4861 caatgtatct tattcgactg tttaaacctg caggcatgca agcttggcgt aatcatggtc
     4921 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4981 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5161 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5221 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
//
