LOCUS       sequence-153098-        3235 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 3235)
  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 3235)
  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-SpyTag-PuroR
FEATURES             Location/Qualifiers
     source          1..3235
                     /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..256
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             266..319
                     /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    943..1064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(954..973)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1008..1027
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1312..1329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1483..2071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1563..1582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2271..3062,3063..3131))
                     /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 gggatccgcc cacatcgtga tggtggacgc
      241 ctacaagccc accaagtccg gaggagaggg cagaggaagt ctgctaacat gcggtgacgt
      301 cgaggagaat cctggcccaa tgactgaata caagcctact gtcaggttgg ctacaagaga
      361 cgacgttcct agagccgtga gaactctggc tgcagccttc gccgactacc ccgccacgag
      421 acacaccgtt gacccagatc ggcatattga gagagtgact gaactgcagg agctgtttct
      481 tacaagagtt ggcctcgaca taggcaaggt gtgggtggcg gacgacggcg ccgccgtggc
      541 cgtctggacc actcccgaat cagttgaggc tggcgccgta ttcgctgaga tcggcccgag
      601 aatggctgag ctcagcggga gtaggctcgc ggcacagcag caaatggagg gactgctggc
      661 accacacagg cccaaagaac ccgcctggtt cctggcaacc gtcggtgtat ctcccgatca
      721 tcaggggaaa ggtctgggct ctgccgtagt gctccctggc gtggaggcag ctgagagagc
      781 aggagtacct gccttcttgg agacctccgc tccaaggaat cttcccttct atgaacggtt
      841 gggcttcacc gtgacagccg acgtggaagt ccccgaaggc ccccgcactt ggtgcatgac
      901 gaggaagcct ggagcgtgag ggcccgcttc gagcagacat gataagatac attgatgagt
      961 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1021 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1081 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1141 tctacaaatg tggtaaaatc gataaagatc cgtcgaccga tgcccttgag agccttcaac
     1201 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     1261 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     1321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2221 tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc
     2281 ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga
     2341 ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca
     2401 atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca
     2461 gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc
     2521 tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc
     2581 attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt
     2641 tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc
     2701 ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg
     2761 gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc
     2821 gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg
     2881 gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg
     2941 aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata
     3001 tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg
     3061 tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt
     3121 tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac
     3181 gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag gccgc
//
