LOCUS       sequence-153102-        3910 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 3910)
  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 3910)
  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-TagGFP2-PuroR
FEATURES             Location/Qualifiers
     source          1..3910
                     /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..931
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     CDS             941..994
                     /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    1618..1739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1629..1648)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1683..1702
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1987..2004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2158..2746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2238..2257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2946..3737,3738..3806))
                     /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 gggatccatg agcgggggcg aggagctgtt
      241 cgccggcatc gtgcccgtgc tgatcgagct ggacggcgac gtgcacggcc acaagttcag
      301 cgtgcgcggc gagggcgagg gcgacgccga ctacggcaag ctggagatca agttcatctg
      361 caccaccggc aagctgcccg tgccctggcc caccctggtg accaccctct gctacggcat
      421 ccagtgcttc gcccgctacc ccgagcacat gaagatgaac gacttcttca agagcgccat
      481 gcccgagggc tacatccagg agcgcaccat ccagttccag gacgacggca agtacaagac
      541 ccgcggcgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcaa
      601 ggacttcaag gaggacggca acatcctggg ccacaagctg gagtacagct tcaacagcca
      661 caacgtgtac atccgccccg acaaggccaa caacggcctg gaggctaact tcaagacccg
      721 ccacaacatc gagggcggcg gcgtgcagct ggccgaccac taccagacca acgtgcccct
      781 gggcgacggc cccgtgctga tccccatcaa ccactacctg agcactcaga ccaagatcag
      841 caaggaccgc aacgaggccc gcgaccacat ggtgctcctg gagtccttca gcgcctgctg
      901 ccacacccac ggcatggacg agctgtacag gtccggagga gagggcagag gaagtctgct
      961 aacatgcggt gacgtcgagg agaatcctgg cccaatgact gaatacaagc ctactgtcag
     1021 gttggctaca agagacgacg ttcctagagc cgtgagaact ctggctgcag ccttcgccga
     1081 ctaccccgcc acgagacaca ccgttgaccc agatcggcat attgagagag tgactgaact
     1141 gcaggagctg tttcttacaa gagttggcct cgacataggc aaggtgtggg tggcggacga
     1201 cggcgccgcc gtggccgtct ggaccactcc cgaatcagtt gaggctggcg ccgtattcgc
     1261 tgagatcggc ccgagaatgg ctgagctcag cgggagtagg ctcgcggcac agcagcaaat
     1321 ggagggactg ctggcaccac acaggcccaa agaacccgcc tggttcctgg caaccgtcgg
     1381 tgtatctccc gatcatcagg ggaaaggtct gggctctgcc gtagtgctcc ctggcgtgga
     1441 ggcagctgag agagcaggag tacctgcctt cttggagacc tccgctccaa ggaatcttcc
     1501 cttctatgaa cggttgggct tcaccgtgac agccgacgtg gaagtccccg aaggcccccg
     1561 cacttggtgc atgacgagga agcctggagc gtgagggccc gcttcgagca gacatgataa
     1621 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1681 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1741 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1801 aaagcaagta aaacctctac aaatgtggta aaatcgataa agatccgtcg accgatgccc
     1861 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     1921 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     1981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2881 aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc
     2941 agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg
     3001 tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca
     3061 ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca
     3121 atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc
     3181 atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc
     3241 cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct
     3301 tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga
     3361 aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg
     3421 tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc
     3481 ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca
     3541 agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa
     3601 gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg
     3661 agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc
     3721 accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt
     3781 gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat
     3841 cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg ggaggtattg
     3901 gacaggccgc
//
