LOCUS       sequence-249714-        3907 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 3907)
  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 3907)
  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-TagRFP-PuroR
FEATURES             Location/Qualifiers
     source          1..3907
                     /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..928
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             938..991
                     /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    1615..1736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1626..1645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1680..1699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1984..2001)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2155..2743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2235..2254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2943..3734,3735..3803))
                     /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 gtgtctaagg gcgaagagct
      241 gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt
      301 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
      361 ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta
      421 cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt
      481 ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc
      541 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt
      601 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccaacac
      661 cgagatgctg taccccgctg acggcggcct ggaaggcaga agcgacatgg ccctgaagct
      721 cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca agaaacccgc
      781 taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa gaatcaagga
      841 ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct
      901 ccctagcaaa ctggggcaca aacttaattc cggaggagag ggcagaggaa gtctgctaac
      961 atgcggtgac gtcgaggaga atcctggccc aatgactgaa tacaagccta ctgtcaggtt
     1021 ggctacaaga gacgacgttc ctagagccgt gagaactctg gctgcagcct tcgccgacta
     1081 ccccgccacg agacacaccg ttgacccaga tcggcatatt gagagagtga ctgaactgca
     1141 ggagctgttt cttacaagag ttggcctcga cataggcaag gtgtgggtgg cggacgacgg
     1201 cgccgccgtg gccgtctgga ccactcccga atcagttgag gctggcgccg tattcgctga
     1261 gatcggcccg agaatggctg agctcagcgg gagtaggctc gcggcacagc agcaaatgga
     1321 gggactgctg gcaccacaca ggcccaaaga acccgcctgg ttcctggcaa ccgtcggtgt
     1381 atctcccgat catcagggga aaggtctggg ctctgccgta gtgctccctg gcgtggaggc
     1441 agctgagaga gcaggagtac ctgccttctt ggagacctcc gctccaagga atcttccctt
     1501 ctatgaacgg ttgggcttca ccgtgacagc cgacgtggaa gtccccgaag gcccccgcac
     1561 ttggtgcatg acgaggaagc ctggagcgtg agggcccgct tcgagcagac atgataagat
     1621 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1681 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1741 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     1801 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataaaga tccgtcgacc gatgcccttg
     1861 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     1921 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
     1981 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2041 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2101 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2161 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2221 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2281 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2341 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2401 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2461 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2521 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2581 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2641 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2701 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2761 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2821 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2881 ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa ccactgcagt
     2941 ggttaccagt gcttgatcag tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc
     3001 atagtggcct gactccccgt cgtgtagatc actacgattc gtgagggctt accatcaggc
     3061 cccagcgcag caatgatgcc gcgagagccg cgttcaccgg cccccgattt gtcagcaatg
     3121 aaccagccag cagggagggc cgagcgaaga agtggtcctg ctactttgtc cgcctccatc
     3181 cagtctatga gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag tagtttccga
     3241 agagttgtgg ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg
     3301 ttcaactctg gttcccagcg gtcaagccgg gtcacatgat cacccatatt atgaagaaat
     3361 gcagtcagct ccttagggcc tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg
     3421 ctcatggtaa tggcagcact acacaattct cttaccgtca tgccatccgt aagatgcttt
     3481 tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc
     3541 tgctcttgcc cggcgtctat acgggacaac accgcgccac atagcagtac tttgaaagtg
     3601 ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc gctattgaga
     3661 tccagttcga tatagcccac tcttgcaccc agttgatctt cagcatcttt tactttcacc
     3721 agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg aatgagtgcg
     3781 acacgaaaat gttggatgct catactcgtc ctttttcaat attattgaag catttatcag
     3841 ggttactagt acgtctctca aggataagta agtaatatta aggtacggga ggtattggac
     3901 aggccgc
//
