LOCUS       sequence-287309-        3719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the In Vitro transcription of DiySpike2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3719)
  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 3719)
  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: pDiySpike2
FEATURES             Location/Qualifiers
     source          1..3719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     442..461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(680..784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     852..870
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(908..930)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1030..1049
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1243..1265
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1257..1274
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1258..1274
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2315..2334)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2316..2334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2377..2393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2377..2393)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2390..2412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2401..2417
                     /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)."
     promoter        complement(join(2425..2431,2432..2449,2450..2455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2470..2491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2608..2625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2779..3367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2859..2878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3538..3719,1..610,611..679))
                     /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"
ORIGIN
        1 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
       61 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      121 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      181 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      241 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      301 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      361 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      421 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      481 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      541 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      601 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      661 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      721 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      781 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
      841 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
      901 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
      961 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     1021 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     1081 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     1141 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     1201 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     1261 aaacgacggc cagtgaattc aagctttttt tttttttttt tttttttttt tttttctgaa
     1321 agtgggtatg gaaaatgccg gattaggacc cagcgtatgt atcaggaatc aacaagtccg
     1381 tatttcttgc gctccgcgag cttaagtact agcagagatc atacatgacg gttctctgcg
     1441 gtcgattaac cacactgctg agcgcgagta tccgctgaat cgaacgtcga taaattcgca
     1501 gtaccggctt ccccgaccgc tgtccacgcg gctgtcagtt agtggcgacc tgggactcta
     1561 cacataagac gatccgtggc tcctgacgac ctagaccacg gtcctttgcg ccacagtcga
     1621 gtaggcccgt cagagaggcc atagactcgc agtgcgggta agggtaacat catgcagaat
     1681 tgtcccggct atgggtccgt cgatgaaagc gaaggcttcc ctagacaaca cgtgctcttg
     1741 agctaaaggc cccgtaaata acgtaagtgg taccagcacg tatcagcgac ccggcattgt
     1801 gaatattcgc cacgacggat gcgcattagc gttccgcatg aatttcctgc gtaacccgac
     1861 cgtcggccgg cactcagtgg cgggtttgta acagaggcgc gcaagggaga gccgcacccc
     1921 gaactccaca ctgccataga actacgagtc gaagcggacg ccactgtaat cttgcttcag
     1981 aaacgggctc taagagcgca tatagcgtgt gagtacgctc gccgccagct cgtcatggtt
     2041 cgagtgcccc cggcgatcag gtagggggat ttagtccggc aggggccggg ggccgttcgc
     2101 tgactactgc cacacagtct cctgcggcta gctcctcgca caccatgacg aacgacctcg
     2161 tgcagcgcgg tctgtcagtt ttcgtcgtcc aagttcatcg gacccgggtg ctttactacc
     2221 tgtatggatt ggcagtagcc ctaggaacta ggcgccccta gatgccgatc cccatgtcta
     2281 agcacctaac ccaatgcatg atcattacct ggctccctat agtgagtcgt attatctaga
     2341 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2401 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2461 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2521 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacc
//
