LOCUS       sequence-153104-        3214 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 3214)
  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 3214)
  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-TC-PuroR
FEATURES             Location/Qualifiers
     source          1..3214
                     /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..235
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             245..298
                     /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    922..1043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(933..952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     987..1006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1291..1308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1462..2050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1542..1561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2250..3041,3042..3110))
                     /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 gggatcctgc tgccccggct gctgctccgg
      241 aggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaat
      301 gactgaatac aagcctactg tcaggttggc tacaagagac gacgttccta gagccgtgag
      361 aactctggct gcagccttcg ccgactaccc cgccacgaga cacaccgttg acccagatcg
      421 gcatattgag agagtgactg aactgcagga gctgtttctt acaagagttg gcctcgacat
      481 aggcaaggtg tgggtggcgg acgacggcgc cgccgtggcc gtctggacca ctcccgaatc
      541 agttgaggct ggcgccgtat tcgctgagat cggcccgaga atggctgagc tcagcgggag
      601 taggctcgcg gcacagcagc aaatggaggg actgctggca ccacacaggc ccaaagaacc
      661 cgcctggttc ctggcaaccg tcggtgtatc tcccgatcat caggggaaag gtctgggctc
      721 tgccgtagtg ctccctggcg tggaggcagc tgagagagca ggagtacctg ccttcttgga
      781 gacctccgct ccaaggaatc ttcccttcta tgaacggttg ggcttcaccg tgacagccga
      841 cgtggaagtc cccgaaggcc cccgcacttg gtgcatgacg aggaagcctg gagcgtgagg
      901 gcccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
      961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1021 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1081 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     1141 ataaagatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     1201 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     1261 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1861 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2221 cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc
     2281 tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact
     2341 acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt
     2401 tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt
     2461 ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta
     2521 agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta
     2581 tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc
     2641 acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc
     2701 agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt
     2761 accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt
     2821 tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc
     2881 gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa
     2941 gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt
     3001 tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa
     3061 aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt
     3121 tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg ataagtaagt
     3181 aatattaagg tacgggaggt attggacagg ccgc
//
