LOCUS       sequence-153101-        3895 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 3895)
  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 3895)
  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-TagBFP-PuroR
FEATURES             Location/Qualifiers
     source          1..3895
                     /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..916
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             926..979
                     /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    1603..1724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1614..1633)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1668..1687
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1972..1989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2143..2731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2223..2242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2931..3722,3723..3791))
                     /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 agcgagctga ttaaggagaa
      241 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc
      301 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
      361 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac
      421 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt
      481 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
      541 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc
      601 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta
      661 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag
      721 ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa
      781 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga
      841 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact
      901 ggggcacaag cttaattccg gaggagaggg cagaggaagt ctgctaacat gcggtgacgt
      961 cgaggagaat cctggcccaa tgactgaata caagcctact gtcaggttgg ctacaagaga
     1021 cgacgttcct agagccgtga gaactctggc tgcagccttc gccgactacc ccgccacgag
     1081 acacaccgtt gacccagatc ggcatattga gagagtgact gaactgcagg agctgtttct
     1141 tacaagagtt ggcctcgaca taggcaaggt gtgggtggcg gacgacggcg ccgccgtggc
     1201 cgtctggacc actcccgaat cagttgaggc tggcgccgta ttcgctgaga tcggcccgag
     1261 aatggctgag ctcagcggga gtaggctcgc ggcacagcag caaatggagg gactgctggc
     1321 accacacagg cccaaagaac ccgcctggtt cctggcaacc gtcggtgtat ctcccgatca
     1381 tcaggggaaa ggtctgggct ctgccgtagt gctccctggc gtggaggcag ctgagagagc
     1441 aggagtacct gccttcttgg agacctccgc tccaaggaat cttcccttct atgaacggtt
     1501 gggcttcacc gtgacagccg acgtggaagt ccccgaaggc ccccgcactt ggtgcatgac
     1561 gaggaagcct ggagcgtgag ggcccgcttc gagcagacat gataagatac attgatgagt
     1621 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1681 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1741 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1801 tctacaaatg tggtaaaatc gataaagatc cgtcgaccga tgcccttgag agccttcaac
     1861 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     1921 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     1981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2881 tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc
     2941 ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga
     3001 ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca
     3061 atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca
     3121 gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc
     3181 tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc
     3241 attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt
     3301 tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc
     3361 ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg
     3421 gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc
     3481 gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg
     3541 gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg
     3601 aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata
     3661 tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg
     3721 tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt
     3781 tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac
     3841 gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag gccgc
//
