LOCUS       sequence-246237-        1810 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  destination vector with N-terminal HA tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1810)
  AUTHORS   Hazelbaker DZ, Beccard A, Angelini G, Mazzucato P, Messana A, Lam D,
            Eggan K, Barrett LE
  TITLE     A multiplexed gRNA piggyBac transposon system facilitates efficient 
            induction of CRISPRi and CRISPRa in human pluripotent stem cells.
  JOURNAL   Sci Rep. 2020 Jan 20;10(1):635. doi: 10.1038/s41598-020-57500-1.
  PUBMED    31959800
REFERENCE   2  (bases 1 to 1810)
  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: "Sci Rep."; 
            date: "2020-01-20"; volume: "10(1):635. doi"; pages: " 
            10.1038/s41598-020-57500-1"
COMMENT     Sequence Label: S2_AscI_HA-gateway_Bsu36I
FEATURES             Location/Qualifiers
     source          1..1810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             64..90
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    91..215
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(240..245,246..263,264..270)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             324..983
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(390..409)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1325..1630
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1509..1528
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1671..1795)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggcgcgccta tcgaaattca ctatagggag acccaagctg gctagttaag cttcgccacc
       61 atgtacccat acgatgttcc agattacgct acaagtttgt acaaaaaagc tgaacgagaa
      121 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
      181 taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct
      241 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccggcgagat
      301 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
      361 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
      421 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
      481 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
      541 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
      601 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
      661 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
      721 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
      781 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
      841 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
      901 atgccgtctg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
      961 atgagtggca gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt
     1021 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     1081 gtatgtcaaa aagaggtgtg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     1141 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     1201 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     1261 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa cagggactgg
     1321 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     1381 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     1441 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     1501 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     1561 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     1621 gggaatataa atgtcaggct ccgttataca cagccagtct gcaggtcgac catagtgact
     1681 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     1741 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtagtaa
     1801 tgaccttagg
//
