LOCUS       sequence-153097-        3310 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 3310)
  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 3310)
  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-SBP-PuroR
FEATURES             Location/Qualifiers
     source          1..3310
                     /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..331
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             341..394
                     /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    1018..1139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1029..1048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1083..1102
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1387..1404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1558..2146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1638..1657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2346..3137,3138..3206))
                     /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 gacgaaaaga ctactggctg
      241 gcgcggagga cacgtcgtcg agggacttgc tggggagctg gaacagcttc gagctaggct
      301 cgagcaccac ccccaggggc aaagagagcc ctccggagga gagggcagag gaagtctgct
      361 aacatgcggt gacgtcgagg agaatcctgg cccaatgact gaatacaagc ctactgtcag
      421 gttggctaca agagacgacg ttcctagagc cgtgagaact ctggctgcag ccttcgccga
      481 ctaccccgcc acgagacaca ccgttgaccc agatcggcat attgagagag tgactgaact
      541 gcaggagctg tttcttacaa gagttggcct cgacataggc aaggtgtggg tggcggacga
      601 cggcgccgcc gtggccgtct ggaccactcc cgaatcagtt gaggctggcg ccgtattcgc
      661 tgagatcggc ccgagaatgg ctgagctcag cgggagtagg ctcgcggcac agcagcaaat
      721 ggagggactg ctggcaccac acaggcccaa agaacccgcc tggttcctgg caaccgtcgg
      781 tgtatctccc gatcatcagg ggaaaggtct gggctctgcc gtagtgctcc ctggcgtgga
      841 ggcagctgag agagcaggag tacctgcctt cttggagacc tccgctccaa ggaatcttcc
      901 cttctatgaa cggttgggct tcaccgtgac agccgacgtg gaagtccccg aaggcccccg
      961 cacttggtgc atgacgagga agcctggagc gtgagggccc gcttcgagca gacatgataa
     1021 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1081 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1141 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1201 aaagcaagta aaacctctac aaatgtggta aaatcgataa agatccgtcg accgatgccc
     1261 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     1321 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     1381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2281 aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc
     2341 agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg
     2401 tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca
     2461 ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca
     2521 atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc
     2581 atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc
     2641 cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct
     2701 tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga
     2761 aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg
     2821 tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc
     2881 ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca
     2941 agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa
     3001 gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg
     3061 agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc
     3121 accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt
     3181 gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat
     3241 cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg ggaggtattg
     3301 gacaggccgc
//
