LOCUS       sequence-287297-        3718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the In Vitro transcription of DiySpike1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3718)
  AUTHORS   Hagemann-Jensen M, Ziegenhain C, Chen P, Ramskold D, Hendriks GJ, 
            Larsson AJM, Faridani OR, Sandberg R
  TITLE     Single-cell RNA counting at allele and isoform resolution using 
            Smart-seq3.
  JOURNAL   Nat Biotechnol. 2020 Jun;38(6):708-714. doi: 
            10.1038/s41587-020-0497-0. Epub 2020 May 4.
  PUBMED    32518404
REFERENCE   2  (bases 1 to 3718)
  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/s41587-020-0497-0"; journalName: "Nat Biotechnol"; date: 
            "2020-06"; volume: "38"; issue: "6"; pages: "708-714"
COMMENT     Sequence Label: pDiySpike1
FEATURES             Location/Qualifiers
     source          1..3718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      272..860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     761..780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1014..1031
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1148..1169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1184..1189,1190..1207,1208..1214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1222..1238
                     /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     1227..1249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1246..1262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1246..1262
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1305..1324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1305..1323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2365..2382)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2365..2381)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2589..2608)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2708..2730
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2768..2786)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2854..2958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2959..3027,3028..3718,1..101)
                     /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(3177..3196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
       61 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      121 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      181 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      241 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      301 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      361 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      421 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      481 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      541 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      601 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      661 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      721 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      781 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      841 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      901 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      961 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1021 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1081 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1141 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1201 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1261 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagataatac gactcactat
     1321 aggggttgcc ttgcaagact agttgcttca taccgcggcg ctgtctaggg ccggggattt
     1381 cgggaagtgt agctcgcttg gcgacctcga tagtgggccc gaccaaaggt cggagccata
     1441 gggtcatact cgaaactggg tcttgaccgc ggttacggcg aggaggggag accacctttc
     1501 ggacctttag cattatagtt acttaatcat gatctgtcgg acgtgggtga gagatgctgt
     1561 gtcgagacaa ctcgcgccga tctgtacaga agctccggac tgcgtatctc cttggatggt
     1621 ggagctggcg gttctcgtga caagaacaca agggcggatc gagctctggt aaagtatgtg
     1681 cactggggac gatcgagccg tcttgccgcg tgacaagcaa actaccaccg taagaagcct
     1741 taaccgagtc aaccccacgg ggtttcctgt taatcgctag cggcatgtct atctcgtcag
     1801 ctagcatgag tcacacggaa cacctgatca tgagggcgcc caaagcaagt acggacgttg
     1861 tatgattaga ctagggaccc gaagctaccg agataccatt tgtgccgacg acattcaacc
     1921 gtatgtggca atctgaggac gcgtgtgtct cattatcttc aagtcgttat taagcggagc
     1981 gctttctcgt tgcggtttaa aagatctccc tcgccgtcgg aatctgggca gtctaacata
     2041 gttatggggg cggagtgatg ggcagaatcc acccacgcac gtctccggac ggaacatacg
     2101 agaggcgtgt gcctgtggtg catgacggta cttttgtgcg gcgcggtatt catgacctat
     2161 gccgcggagc gattcacctg acccgactgt cgataggaga ccggagcgac tttttgttac
     2221 acataagcga tgactgtgaa ccgtgtgagg gggtcctcct attaggttac aacattatga
     2281 tactacgagc ggtcggccaa gttgagttcc tgtaccaaaa ccaaaaaaaa aaaaaaaaaa
     2341 aaaaaaaaaa aaaagcttga attcactggc cgtcgtttta caactcgtga ctgggaaaac
     2401 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2461 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2521 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     2581 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     2641 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2701 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     2761 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2821 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2881 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2941 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3001 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3061 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3121 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3181 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3241 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3301 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3361 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3421 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3481 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3541 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3601 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3661 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccct
//
