LOCUS       sequence-153099-        3220 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 3220)
  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 3220)
  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-StrepTag II-PuroR
FEATURES             Location/Qualifiers
     source          1..3220
                     /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..241
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             251..304
                     /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    928..1049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(939..958)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     993..1012
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1297..1314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1468..2056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1548..1567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2256..3047,3048..3116))
                     /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 gggatcctgg agccaccccc agttcgagaa
      241 gtccggagga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
      301 cccaatgact gaatacaagc ctactgtcag gttggctaca agagacgacg ttcctagagc
      361 cgtgagaact ctggctgcag ccttcgccga ctaccccgcc acgagacaca ccgttgaccc
      421 agatcggcat attgagagag tgactgaact gcaggagctg tttcttacaa gagttggcct
      481 cgacataggc aaggtgtggg tggcggacga cggcgccgcc gtggccgtct ggaccactcc
      541 cgaatcagtt gaggctggcg ccgtattcgc tgagatcggc ccgagaatgg ctgagctcag
      601 cgggagtagg ctcgcggcac agcagcaaat ggagggactg ctggcaccac acaggcccaa
      661 agaacccgcc tggttcctgg caaccgtcgg tgtatctccc gatcatcagg ggaaaggtct
      721 gggctctgcc gtagtgctcc ctggcgtgga ggcagctgag agagcaggag tacctgcctt
      781 cttggagacc tccgctccaa ggaatcttcc cttctatgaa cggttgggct tcaccgtgac
      841 agccgacgtg gaagtccccg aaggcccccg cacttggtgc atgacgagga agcctggagc
      901 gtgagggccc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
      961 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1021 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1081 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1141 aaatcgataa agatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     1201 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     1261 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2221 tgacagcggc cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca
     2281 ccgatctcag cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag
     2341 atcactacga ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag
     2401 ccgcgttcac cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga
     2461 agaagtggtc ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct
     2521 agagtaagaa gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc
     2581 gtggtatcac gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc
     2641 cgggtcacat gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc
     2701 gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat
     2761 tctcttaccg tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag
     2821 tcgttttgtg agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac
     2881 aacaccgcgc cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg
     2941 cggaaagact caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca
     3001 cccagttgat cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc
     3061 aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc
     3121 gtcctttttc aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa
     3181 gtaagtaata ttaaggtacg ggaggtattg gacaggccgc
//
