LOCUS       sequence-153092-        4081 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 4081)
  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 4081)
  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-HaloTag-PuroR
FEATURES             Location/Qualifiers
     source          1..4081
                     /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..1102
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             1112..1165
                     /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    1789..1910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1800..1819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1854..1873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2158..2175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2329..2917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2409..2428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3117..3908,3909..3977))
                     /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 gggatccgaa atcggtactg gctttccatt
      241 cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg
      301 cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa
      361 catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg
      421 caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc
      481 cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc
      541 tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga
      601 gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca
      661 ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga
      721 gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg
      781 cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc
      841 aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca
      901 ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc
      961 cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg
     1021 tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct
     1081 gtctactctg gagatttccg gttccggagg agagggcaga ggaagtctgc taacatgcgg
     1141 tgacgtcgag gagaatcctg gcccaatgac tgaatacaag cctactgtca ggttggctac
     1201 aagagacgac gttcctagag ccgtgagaac tctggctgca gccttcgccg actaccccgc
     1261 cacgagacac accgttgacc cagatcggca tattgagaga gtgactgaac tgcaggagct
     1321 gtttcttaca agagttggcc tcgacatagg caaggtgtgg gtggcggacg acggcgccgc
     1381 cgtggccgtc tggaccactc ccgaatcagt tgaggctggc gccgtattcg ctgagatcgg
     1441 cccgagaatg gctgagctca gcgggagtag gctcgcggca cagcagcaaa tggagggact
     1501 gctggcacca cacaggccca aagaacccgc ctggttcctg gcaaccgtcg gtgtatctcc
     1561 cgatcatcag gggaaaggtc tgggctctgc cgtagtgctc cctggcgtgg aggcagctga
     1621 gagagcagga gtacctgcct tcttggagac ctccgctcca aggaatcttc ccttctatga
     1681 acggttgggc ttcaccgtga cagccgacgt ggaagtcccc gaaggccccc gcacttggtg
     1741 catgacgagg aagcctggag cgtgagggcc cgcttcgagc agacatgata agatacattg
     1801 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1861 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1921 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1981 aaaacctcta caaatgtggt aaaatcgata aagatccgtc gaccgatgcc cttgagagcc
     2041 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2101 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     2161 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2221 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2281 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2521 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2701 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2761 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2821 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3061 tatatatgag taaacttggt ctgacagcgg ccgcaaatgc taaaccactg cagtggttac
     3121 cagtgcttga tcagtgaggc accgatctca gcgatctgcc tatttcgttc gtccatagtg
     3181 gcctgactcc ccgtcgtgta gatcactacg attcgtgagg gcttaccatc aggccccagc
     3241 gcagcaatga tgccgcgaga gccgcgttca ccggcccccg atttgtcagc aatgaaccag
     3301 ccagcaggga gggccgagcg aagaagtggt cctgctactt tgtccgcctc catccagtct
     3361 atgagctgct gtcgtgatgc tagagtaaga agttcgccag tgagtagttt ccgaagagtt
     3421 gtggccattg ctactggcat cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac
     3481 tctggttccc agcggtcaag ccgggtcaca tgatcaccca tattatgaag aaatgcagtc
     3541 agctccttag ggcctccgat cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg
     3601 gtaatggcag cactacacaa ttctcttacc gtcatgccat ccgtaagatg cttttccgtg
     3661 accggcgagt actcaaccaa gtcgttttgt gagtagtgta tacggcgacc aagctgctct
     3721 tgcccggcgt ctatacggga caacaccgcg ccacatagca gtactttgaa agtgctcatc
     3781 atcgggaatc gttcttcggg gcggaaagac tcaaggatct tgccgctatt gagatccagt
     3841 tcgatatagc ccactcttgc acccagttga tcttcagcat cttttacttt caccagcgtt
     3901 tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga agggaatgag tgcgacacga
     3961 aaatgttgga tgctcatact cgtccttttt caatattatt gaagcattta tcagggttac
     4021 tagtacgtct ctcaaggata agtaagtaat attaaggtac gggaggtatt ggacaggccg
     4081 c
//
