LOCUS       sequence-153095-        3550 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 3550)
  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 3550)
  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-nTEV-PuroR
FEATURES             Location/Qualifiers
     source          1..3550
                     /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             581..634
                     /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    1258..1379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1269..1288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1323..1342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1627..1644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1798..2386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1878..1897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2586..3377,3378..3446))
                     /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 gggatccggc gagagcctgt tcaagggacc
      241 cagagactac aaccccatca gcagcaccat ctgccacctg accaacgaga gcgacggcca
      301 caccaccagc ctgtacggca tcggcttcgg ccccttcatc atcaccaaca agcacctgtt
      361 cagaagaaac aacggcaccc tgctggtgca gagcctgcac ggcgtgttca aggtgaagaa
      421 caccaccacc ctgcagcagc acctgatcga cggcagagac atgatcatca tcagaatgcc
      481 caaggacttc ccccccttcc cccagaagct gaagttcaga gagccccaga gagaggagag
      541 aatctgcctg gtgaccacca acttccagac ctccggagga gagggcagag gaagtctgct
      601 aacatgcggt gacgtcgagg agaatcctgg cccaatgact gaatacaagc ctactgtcag
      661 gttggctaca agagacgacg ttcctagagc cgtgagaact ctggctgcag ccttcgccga
      721 ctaccccgcc acgagacaca ccgttgaccc agatcggcat attgagagag tgactgaact
      781 gcaggagctg tttcttacaa gagttggcct cgacataggc aaggtgtggg tggcggacga
      841 cggcgccgcc gtggccgtct ggaccactcc cgaatcagtt gaggctggcg ccgtattcgc
      901 tgagatcggc ccgagaatgg ctgagctcag cgggagtagg ctcgcggcac agcagcaaat
      961 ggagggactg ctggcaccac acaggcccaa agaacccgcc tggttcctgg caaccgtcgg
     1021 tgtatctccc gatcatcagg ggaaaggtct gggctctgcc gtagtgctcc ctggcgtgga
     1081 ggcagctgag agagcaggag tacctgcctt cttggagacc tccgctccaa ggaatcttcc
     1141 cttctatgaa cggttgggct tcaccgtgac agccgacgtg gaagtccccg aaggcccccg
     1201 cacttggtgc atgacgagga agcctggagc gtgagggccc gcttcgagca gacatgataa
     1261 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1321 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1381 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1441 aaagcaagta aaacctctac aaatgtggta aaatcgataa agatccgtcg accgatgccc
     1501 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     1561 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     1621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2221 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2521 aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc
     2581 agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg
     2641 tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca
     2701 ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca
     2761 atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc
     2821 atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc
     2881 cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct
     2941 tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga
     3001 aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg
     3061 tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc
     3121 ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca
     3181 agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa
     3241 gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg
     3301 agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc
     3361 accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt
     3421 gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat
     3481 cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg ggaggtattg
     3541 gacaggccgc
//
