LOCUS       sequence-153090-        3568 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 3568)
  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 3568)
  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-cTEV-PuroR
FEATURES             Location/Qualifiers
     source          1..3568
                     /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             599..652
                     /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    1276..1397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1287..1306)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1341..1360
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1645..1662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1816..2404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1896..1915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2604..3395,3396..3464))
                     /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 gggatccaag agcatgagca gcatggtgag
      241 cgacaccagc tgcaccttcc ccagcagcga cggcatcttc tggaagcact ggattcagac
      301 caaggacggc cagtgcggca gccccctggt gagcaccaga gacggcttca tcgtgggcat
      361 ccacagcgcc agcaacttca ccaacaccaa caactacttc accagcgtgc ccaagaactt
      421 catggagctg ctgaccaacc aggaggccca gcagtgggtg agcggctgga gactgaacgc
      481 cgacagcgtg ctgtggggcg gccacaaggt gttcatgagc aagcccgagg agcccttcca
      541 gcccgtgaag gaggccaccc agctgatgaa cgagctggtg tacagccagt ccggaggaga
      601 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc caatgactga
      661 atacaagcct actgtcaggt tggctacaag agacgacgtt cctagagccg tgagaactct
      721 ggctgcagcc ttcgccgact accccgccac gagacacacc gttgacccag atcggcatat
      781 tgagagagtg actgaactgc aggagctgtt tcttacaaga gttggcctcg acataggcaa
      841 ggtgtgggtg gcggacgacg gcgccgccgt ggccgtctgg accactcccg aatcagttga
      901 ggctggcgcc gtattcgctg agatcggccc gagaatggct gagctcagcg ggagtaggct
      961 cgcggcacag cagcaaatgg agggactgct ggcaccacac aggcccaaag aacccgcctg
     1021 gttcctggca accgtcggtg tatctcccga tcatcagggg aaaggtctgg gctctgccgt
     1081 agtgctccct ggcgtggagg cagctgagag agcaggagta cctgccttct tggagacctc
     1141 cgctccaagg aatcttccct tctatgaacg gttgggcttc accgtgacag ccgacgtgga
     1201 agtccccgaa ggcccccgca cttggtgcat gacgaggaag cctggagcgt gagggcccgc
     1261 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1321 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1381 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1441 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataaag
     1501 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1561 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     1621 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1681 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1741 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1801 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1861 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1921 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1981 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2041 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2101 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2161 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2221 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2281 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2341 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2401 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2461 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2521 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagcggccg
     2581 caaatgctaa accactgcag tggttaccag tgcttgatca gtgaggcacc gatctcagcg
     2641 atctgcctat ttcgttcgtc catagtggcc tgactccccg tcgtgtagat cactacgatt
     2701 cgtgagggct taccatcagg ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg
     2761 gcccccgatt tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag aagtggtcct
     2821 gctactttgt ccgcctccat ccagtctatg agctgctgtc gtgatgctag agtaagaagt
     2881 tcgccagtga gtagtttccg aagagttgtg gccattgcta ctggcatcgt ggtatcacgc
     2941 tcgtcgttcg gtatggcttc gttcaactct ggttcccagc ggtcaagccg ggtcacatga
     3001 tcacccatat tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt
     3061 aagttggccg cggtgttgtc gctcatggta atggcagcac tacacaattc tcttaccgtc
     3121 atgccatccg taagatgctt ttccgtgacc ggcgagtact caaccaagtc gttttgtgag
     3181 tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca
     3241 catagcagta ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg gaaagactca
     3301 aggatcttgc cgctattgag atccagttcg atatagccca ctcttgcacc cagttgatct
     3361 tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc
     3421 gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt cctttttcaa
     3481 tattattgaa gcatttatca gggttactag tacgtctctc aaggataagt aagtaatatt
     3541 aaggtacggg aggtattgga caggccgc
//
