LOCUS       sequence-287360-        3620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the In Vitro transcription of DiySpike1d4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3620)
  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 3620)
  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: pDiySpike1d4
FEATURES             Location/Qualifiers
     source          1..3620
                     /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(2275..2297)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2491..2510)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2610..2632
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2670..2688)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2756..2860
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2861..2929,2930..3620,1..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(3079..3098)
                     /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 ttgcaagagt 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 ccataagcga tgactgtgaa ccgtgtgagg gggtcctcct
     2161 attaggttac aacattatga tactacgagc ggtcggccaa gttgagttcc tgtaccaaaa
     2221 ccaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaagcttga attcactggg ccgtcgtttt
     2281 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2341 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2401 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2461 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2521 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2581 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2641 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2701 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2761 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2821 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2881 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2941 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3001 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3061 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3121 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3181 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3241 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3301 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3361 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3421 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3481 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3541 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3601 ggggccagat ggtaagccct
//
