LOCUS       sequence-474378-        5847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TRF1-PYL1 for recruiting RNA to the telomere via 
            CRISPR-TO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5847)
  AUTHORS   Han M, Fu ML, Zhu Y, Choi AA, Li E, Bezney J, Cai S, Miles L, Ma Y, 
            Qi LS
  TITLE     Programmable control of spatial transcriptome in live cells and 
            neurons.
  JOURNAL   Nature. 2025 Jul;643(8070):241-251. doi: 10.1038/s41586-025-09020-z.
            Epub 2025 May 21.
  PUBMED    40399675
REFERENCE   2  (bases 1 to 5847)
  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/s41586-025-09020-z"; journalName: "Nature"; date: 
            "2025-07"; volume: "643"; issue: "8070"; pages: "241-251"
COMMENT     Sequence Label: pSLQ14566
FEATURES             Location/Qualifiers
     source          1..5847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        771..1074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1075..1278
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1229..1249
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2683..3378
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    4073..4194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4110..4129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4164..4183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4201..4656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4288..4307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4498..4519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4817..4885,4886..5677)
                     /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     complement(5035..5054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
      661 cccctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
      721 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      781 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      841 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      901 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      961 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1021 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1081 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1141 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1201 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1261 gtctatataa gcagagctga tatcaagctt gcggtacgac tgagtcgcgg ccgcatggcg
     1321 gaggatgttt cctcagcggc cctgagcccg cggggctgtg cggatggtag ggatgccgac
     1381 cctactgagg agcagatggc agaaacagag agaaacgacg aggagcagtt cgaatgccag
     1441 gaactgctcg agtgccaggt gcaggtgggg gcccccgagg aggaggagga ggaggaggag
     1501 gacgcgggcc tggtggccga ggccgaggcc gtggctgccg gctggatgct cgatttcctc
     1561 tgcctctctc tttgccgagc tttccgcgac ggccgctccg aggacttccg caggacccgc
     1621 aacagcgcag aggctattat tcatggacta tccagtctaa cagcttgcca gttgagaacg
     1681 atatacatat gtcagttttt gacaagaatt gcagcaggaa aaacccttga tgcacagttt
     1741 gaaaatgatg aacgaattac acccttggaa tcagccctga tgatttgggg ttcaattgaa
     1801 aaggaacatg acaaacttca tgaagaaata cagaatttaa ttaaaattca ggctatagct
     1861 gtttgtatgg aaaatggcaa ctttaaagaa gcagaagaag tctttgaaag aatatttggt
     1921 gatccaaatt ctcatatgcc tttcaaaagc aaattgctta tgataatctc tcagaaagat
     1981 acatttcatt ccttttttca acacttcagc tacaaccaca tgatggagaa aattaagagt
     2041 tatgtgaatt atgtgctaag tgaaaaatca tcaacctttc taatgaaggc agcggcaaaa
     2101 gtagtagaaa gcaaaaggac aagaacaata acttctcaag ataaacctag tggtaatgat
     2161 gttgaaatgg aaactgaagc taatttggat acaagaaaaa gtgttagtga caaacagtct
     2221 gcggtaactg aatcctcaga gggtacagta tccttattga ggtctcacaa gaatcttttc
     2281 ttatctaagt tgcaacatgg aacccagcaa caagacctta ataagaaaga aagaagagta
     2341 ggaactcctc aaagtacaaa aaagaaaaaa gaaagcagaa gagccactga aagcagaata
     2401 cctgtttcaa agagtcagcc ggtaactcct gaaaaacatc gagctagaaa aagacaggca
     2461 tggctttggg aagaagacaa gaatttgaga tctggcgtga ggaaatatgg agagggaaac
     2521 tggtctaaaa tactgttgca ttataaattc aacaaccgga caagtgtcat gttaaaagac
     2581 agatggagga ccatgaagaa actaaaactg atttcctcag acagcgaaga ctgcggcgga
     2641 ccgggatcag gaagcgtgga tccaggtgga ggtggaagcg gtagcgagct gattaaggag
     2701 aacatgcaca tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca
     2761 tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag
     2821 ggcggccctc tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag
     2881 accttcatca accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc
     2941 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac
     3001 accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca
     3061 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg
     3121 taccccgctg acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg
     3181 agccatctga tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc
     3241 aagatgcctg gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac
     3301 gagacctacg tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa
     3361 ctggggcaca agcttaatgg atcaggaagc ggtgggggcg cgccaactca agacgaattc
     3421 acccaactct cccaatcaat cgccgagttc cacacgtacc aactcggtaa cggccgttgc
     3481 tcatctctcc tagctcagcg aatccacgcg ccgccggaaa cagtatggtc cgtggtgaga
     3541 cgtttcgata ggccacagat ttacaaacac ttcatcaaaa gctgtaacgt gagtgaagat
     3601 ttcgagatgc gagtgggatg cacgcgcgac gtgaacgtga taagtggatt accggcgaat
     3661 acgtctcgag agagattaga tctgttggac gatgatcgga gagtgactgg gtttagtata
     3721 accggtggtg aacataggct gaggaattat aaatcggtta cgacggttca tagatttgag
     3781 aaagaagaag aagaagaaag gatctggacc gttgttttgg aatcttatgt tgttgatgta
     3841 ccggaaggta attcggagga agatacgaga ttgtttgctg atacggttat tagattgaat
     3901 cttcagaaac ttgcttcgat cactgaagct atgaactgac ctgcaggggt accggccgcg
     3961 acactagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     4021 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     4081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     4201 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4261 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4321 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4381 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4441 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4501 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4561 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4621 acgcgaattt taacaaaata ttaacgctta caatttcctc tagataataa tggtttctta
     4681 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4861 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4921 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4981 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5041 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5101 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5161 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5221 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5281 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5341 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5401 actacttact ctagcttccc ggcaacaact gatagactgg atggaggcgg ataaagttgc
     5461 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5521 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5581 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5641 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5701 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5761 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5821 ccccgtagaa aagatcaaag gatcttc
//
