LOCUS       sequence-178474-        4098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for HA-tag embedded into human CD52 with short 
            polyA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4098)
  AUTHORS   Zotova A, Pichugin A, Atemasova A, Knyazhanskaya E, Lopatukhina E, 
            Mitkin N, Holmuhamedov E, Gottikh M, Kuprash D, Filatov A, Mazurov D
  TITLE     Isolation of gene-edited cells via knock-in of short 
            glycophosphatidylinositol-anchored epitope tags.
  JOURNAL   Sci Rep. 2019 Feb 28;9(1):3132. doi: 10.1038/s41598-019-40219-z.
  PUBMED    30816313
REFERENCE   2  (bases 1 to 4098)
  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: "2019-02-28"; volume: "9(1):3132. doi"; pages: " 
            10.1038/s41598-019-40219-z"
COMMENT     Sequence Label: pJet-CMV-CD5HA2-bglpA
FEATURES             Location/Qualifiers
     source          1..4098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..20)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        88..192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(193..261,262..1053)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(411..430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1224..1812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1713..1732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1966..1983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2103..2124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2139..2144,2145..2162,2163..2169)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     enhancer        2684..3063
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3064..3265
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3216..3236
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3262..3286
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3565..3591
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3672..3720
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3775..3794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3776..3794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
       61 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      121 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      181 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      241 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      301 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      361 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      421 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      481 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      541 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      601 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      661 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      721 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
      781 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
      841 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      901 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      961 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1021 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1081 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1141 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1921 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1981 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2041 taatgcagct ggcacgacag gtttcccgac tggaaagcaa ttggcagtga gcgcaacgca
     2101 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     2161 cgtataatgt gtggaattat gagcggataa taatttcaca caggaggttt aaactttaaa
     2221 catgtcaaaa gagacgtctt ttgttaagaa tgctgaggaa cttgcaaagc aaaaaatgga
     2281 tgctattaac cctgaacttt cttcaaaatt taaattttta ataaaattcc tgtctcagtt
     2341 tcctgaagct tgctctaaac ctcgttcaaa aaaaatgcag aataaagttg gtcaagagga
     2401 acatattgaa tatttagctc gtagttttca tgagagtcga ttgccaagaa aacccacgcc
     2461 acctacaacg gttcctgatg aggtggttag catagttctt aatataagtt ttaatataca
     2521 gcctgaaaat cttgagagaa taaaagaaga acatcgattt tccatggcag ctgagaatat
     2581 tgtaggagat cttctagaaa gatccccgcg gccattgcat acgttgtatc catatcataa
     2641 tatgtacatt tatattggct catgtccaca ttaccgccat gttgacattg attattgact
     2701 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     2761 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2821 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2881 tgggtggact atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     2941 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     3001 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     3061 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggat
     3121 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     3181 acttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3241 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     3301 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccggga
     3361 acggtgcatt ggaacgcgga ttccccgtgc caagagtgac gtaagtaccg cctatagagt
     3421 ctataggccc acccccttgg cttcttatgc gacggatctg cctgcaggtc gaccaccatg
     3481 aagcgcttcc tcttcctcct actcaccatc agcctcctgg ttatggtaca gatacaaact
     3541 ggactctcag gacaaaacga cacctaccct tacgacgtgc ccgactacgc ctcagcatcc
     3601 agcaacataa gcggaggcat tttccttttc ttcgtggcca atgccataat ccacctcttc
     3661 tgcttcagtt gaataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     3721 gtaccgcatc ttgctgaaaa actcgagcca tccggaagat ctggcggccg ctctccctat
     3781 agtgagtcgt attacgccgg atggatatgg tgttcaggca caagtgttaa agcagttgat
     3841 tttattcact atgatgaaaa aaacaatgaa tggaacctgc tccaagttaa aaatagagat
     3901 aataccgaaa actcatcgag tagtaagatt agagataata caacaataaa aaaatggttt
     3961 agaacttact cacagcgtga tgctactaat tgggacaatt ttccagatga agtatcatct
     4021 aagaatttaa atgaagaaga cttcagagct tttgttaaaa attatttggc aaaaataata
     4081 taattcggct gcaggggc
//
