LOCUS       sequence-287364-        3619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the In Vitro transcription of DiySpike2d3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3619)
  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 3619)
  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: pDiySpike2d3
FEATURES             Location/Qualifiers
     source          1..3619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      353..941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     842..861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1095..1112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1229..1250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1265..1270,1271..1288,1289..1295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1319
                     /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     1308..1330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1327..1343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1327..1343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1386..1405
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1386..1404
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2346..2363)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2346..2362)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2355..2377)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2571..2590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2690..2712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2750..2768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2836..2940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2941..3009,3010..3619,1..182)
                     /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(3159..3178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
       61 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      481 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1021 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1081 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1201 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1261 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1321 ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact
     1381 ctagataata cgactcacta tagggagcca ggtaatgatc atgcattggg ttaggtggtt
     1441 agacatgggg atcggcatct aggggcgcct agttcctagg gctactgcca atccatacag
     1501 gtagtaaagc acccgggtcc gatgaacttg gacgacgaaa actgacagac cgcgctgcac
     1561 gaggtcgttc gtcatggtgt gcgaggagct agccgcagga gactgtgtgg cagtagtcag
     1621 cgaacggccc ccggcccctg ccggactaaa tccccctacc tgatcgccgg gggcactcga
     1681 accatgacga gctggcggcg agcgtactca cacgctatat gcgctcttag agcccgtttc
     1741 tgaagcaaga ttacagtggc gtccgcttcg actcgtagtt ctatggcagt gtggagttcg
     1801 gggtgcggct ctcccttgcg cgcctctgtt acaaacccgc cactgagtgc cggccgacgg
     1861 tcgggttacg caggaaattc atgcggaacg ctaatgcgca tccgtcgtgg cgaatattca
     1921 caatgccggg tcgctgatac gtgctggtac cacttacgtt atttacgggg cctttagctc
     1981 aagagcacgt gttgtctagg gaagccttcg ctaaaggacc gtggtctagg tcgtcaggag
     2041 ccacggatcg tcttatgtgt agagtcccag gtcgccacta actgacagcc gcgtggacag
     2101 cggtcgggga agccggtact gcgaatttat cgacgttcga ttcagcggat actcgcgctc
     2161 agcagtgtgg ttaatcgacc gcagagaacc gtcatgtatg atctctgcta gtacttaagc
     2221 tcgcggagcg caagaaatac ggacttgttg attcctgata catacgctgg gtcctaatcc
     2281 ggcattttcc atacccactt tcaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaagcttg
     2341 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2401 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2461 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     2521 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     2581 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     2641 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2701 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     2761 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2821 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2881 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2941 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3001 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3061 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3121 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3181 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3241 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3301 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3361 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3421 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3481 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3541 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3601 tcggcccttc cggctggct
//
