LOCUS       sequence-153093-        3226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPaint gene tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3226)
  AUTHORS   Schmid-Burgk JL, Honing K, Ebert TS, Hornung V
  TITLE     CRISPaint allows modular base-specific gene tagging using a 
            ligase-4-dependent mechanism.
  JOURNAL   Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338.
  PUBMED    27465542
REFERENCE   2  (bases 1 to 3226)
  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"; journalName: "Nat 
            Commun."; date: "2016-07-28"; volume: "7:12338. doi"; pages: " 
            10.1038/ncomms12338"
COMMENT     Sequence Label: pCRISPaint-myc-PuroR
FEATURES             Location/Qualifiers
     source          1..3226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             218..247
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             257..310
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    934..1055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(945..964)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     999..1018
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1303..1320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1474..2062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1554..1573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2262..3053,3054..3122))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctcttaatga tagctagcgg gccagtaccc
      181 aaaaagcggg gggtctggtg gcagtggagg gggatccgag cagaagctga tcagcgagga
      241 ggacctgtcc ggaggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa
      301 tcctggccca atgactgaat acaagcctac tgtcaggttg gctacaagag acgacgttcc
      361 tagagccgtg agaactctgg ctgcagcctt cgccgactac cccgccacga gacacaccgt
      421 tgacccagat cggcatattg agagagtgac tgaactgcag gagctgtttc ttacaagagt
      481 tggcctcgac ataggcaagg tgtgggtggc ggacgacggc gccgccgtgg ccgtctggac
      541 cactcccgaa tcagttgagg ctggcgccgt attcgctgag atcggcccga gaatggctga
      601 gctcagcggg agtaggctcg cggcacagca gcaaatggag ggactgctgg caccacacag
      661 gcccaaagaa cccgcctggt tcctggcaac cgtcggtgta tctcccgatc atcaggggaa
      721 aggtctgggc tctgccgtag tgctccctgg cgtggaggca gctgagagag caggagtacc
      781 tgccttcttg gagacctccg ctccaaggaa tcttcccttc tatgaacggt tgggcttcac
      841 cgtgacagcc gacgtggaag tccccgaagg cccccgcact tggtgcatga cgaggaagcc
      901 tggagcgtga gggcccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
      961 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1021 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1081 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1141 gtggtaaaat cgataaagat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc
     1201 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     1261 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
     1321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2101 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2161 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2221 ttggtctgac agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt
     2281 gaggcaccga tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc
     2341 gtgtagatca ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg
     2401 cgagagccgc gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc
     2461 gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt
     2521 gatgctagag taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact
     2581 ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg
     2641 tcaagccggg tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct
     2701 ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta
     2761 cacaattctc ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca
     2821 accaagtcgt tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata
     2881 cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct
     2941 tcggggcgga aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact
     3001 cttgcaccca gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa
     3061 acaggcaagc aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc
     3121 atactcgtcc tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa
     3181 ggataagtaa gtaatattaa ggtacgggag gtattggaca ggccgc
//
