LOCUS       sequence-474373-        2969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses gG2 for pertubing endogenous human GAPDH mRNA via 
            CRISPR-TO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2969)
  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 2969)
  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: pSLQ14902
FEATURES             Location/Qualifiers
     source          1..2969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     229..248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     482..499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    616..637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(652..657,658..675,676..682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    690..706
                     /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)."
     primer_bind     695..717
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     714..730
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     714..730
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        744..992
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     744..764
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     915..934
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1083..1100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1083..1099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1092..1114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1308..1327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1487..1505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1573..1677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1678..1746,1747..2538)
                     /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(1896..1915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2709..2969,1..328)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
       61 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      121 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      181 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      241 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      301 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      361 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      421 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      481 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      541 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      601 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      661 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      721 cagctatgac catgattacg ccagagggcc tatttcccat gattccttca tatttgcata
      781 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
      841 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
      901 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
      961 ctttatatat cttgtggaaa ggacgaaaca ccgtctacat ggcaactgtg aggaggggag
     1021 atgttgtgga aggtccagtt ttgaggggct attacaactt ttttggtacc gagctcgaat
     1081 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1141 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1201 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     1261 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     1321 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1381 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctacgg gagctgcatg
     1441 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcaagac gaaagggcct cgtgatacgc
     1501 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2941 gtggctgctg ccagtggcga taagtcgtg
//
