LOCUS       sequence-287342-        3719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the In Vitro transcription of DiySpike3.
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: pDiySpike3
FEATURES             Location/Qualifiers
     source          1..3719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      305..893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     794..813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1047..1064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1181..1202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1217..1222,1223..1240,1241..1247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1255..1271
                     /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     1260..1282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1279..1295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1279..1295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1338..1356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2398..2415)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2398..2414)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2407..2429)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2623..2642)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2742..2764
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2802..2820)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2888..2992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2993..3061,3062..3719,1..134)
                     /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(3211..3230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
       61 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      121 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      181 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      241 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      301 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      361 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      421 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      481 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      541 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      601 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      661 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      721 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      781 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      841 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      901 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      961 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1021 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1081 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1141 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     1201 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     1261 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc
     1321 tgcaggtcga ctctagataa tacgactcac tataggcttc tacttcccag ccacaaacga
     1381 gaagggtgag catattactc aaaatgtttt ctatcccccc ctaggtggaa cttatatttc
     1441 gaattaacga caaaccggga cggctcaccc ctccatttcg atacccaccg ccgagaggcc
     1501 gacgtatgcg atggctagtt ccgcccacag attataatac caagtcgcgc gagggtacct
     1561 tgattttaaa tgttcctcct ttacgttatg tgtgcggcca caccgtgtcc gtgtttcggt
     1621 cacactttag tgctatgagt aacgtacaca gtcgcgcagt gcacgtatag atggagccct
     1681 tatgttgtta aagaagtgca gtacgcttga tcggtagagc gacaactccc cgagttaagt
     1741 ggcgtcgacc gacgagttac cgccgatggg taaaaatctt tacaagcact tccgaatatt
     1801 aggataatgc gctctttggc aaaataaatg tcggtgctcc gagtggctac tagccttccc
     1861 gaagagtggt gaactagatt gtgcttaatg cacgacctgc ttaggtcact agtatcgtcc
     1921 gacctggcgg gtttctcata ccagtcggcg tacgctgact agtcactcaa agagtagtag
     1981 ggtaactctc gaggcaataa ggtatttgtg ccaggtgcat cctcggtggg actatcggtt
     2041 gccactaccc atagtgatgt cgatgttccg ttagcactgt tcgaaacgtt tcttccgggt
     2101 atcgccaacc attggacgga caataaactt cgcaagagga agtggtggtg tcttagtgca
     2161 caagggccga tattgatgga actcagggtc ggtgggcctc tgcgcagatt gtcaagtttc
     2221 ttccctattc ggtaactgga aaagcacgca ttcagcttga caaatctcgt attgaataca
     2281 ttcgttagca ctagacgtca taatgctggg tgctaggtaa attacttaac tagtgtgacg
     2341 acctgatcag agtctaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaagct tgaattcact
     2401 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2461 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2521 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     2581 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     2641 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2701 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2761 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     2821 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2881 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2941 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3001 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3061 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3121 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3181 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3241 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3301 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3361 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3421 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3481 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3541 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3601 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3661 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggta
//
