LOCUS       sequence-183222-        4101 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 4101)
  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 4101)
  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..4101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    110..131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(146..151,152..169,170..176)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     enhancer        692..1071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1072..1275
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1226..1246
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1272..1296
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1575..1601
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1682..1730
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1785..1804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1786..1804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2110..2128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2196..2300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2301..2369,2370..3161)
                     /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(2519..2538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3332..3920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3821..3840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4074..4091
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcaattg gcagtgagcg caacgcaatt aatgtgagtt
      121 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
      181 gaattatgag cggataataa tttcacacag gaggtttaaa ctttaaacat gtcaaaagag
      241 acgtcttttg ttaagaatgc tgaggaactt gcaaagcaaa aaatggatgc tattaaccct
      301 gaactttctt caaaatttaa atttttaata aaattcctgt ctcagtttcc tgaagcttgc
      361 tctaaacctc gttcaaaaaa aatgcagaat aaagttggtc aagaggaaca tattgaatat
      421 ttagctcgta gttttcatga gagtcgattg ccaagaaaac ccacgccacc tacaacggtt
      481 cctgatgagg tggttagcat agttcttaat ataagtttta atatacagcc tgaaaatctt
      541 gagagaataa aagaagaaca tcgattttcc atggcagctg agaatattgt aggagatctt
      601 ctagaaagat ccccgcggcc attgcatacg ttgtatccat atcataatat gtacatttat
      661 attggctcat gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag
      721 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      781 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      841 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      901 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      961 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1021 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1081 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1141 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1201 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1261 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
     1321 tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccggga acggtgcatt
     1381 ggaacgcgga ttccccgtgc caagagtgac gtaagtaccg cctatagagt ctataggccc
     1441 acccccttgg cttcttatgc gacggatctg cctgcaggtc gaccaccatg aagcgcttcc
     1501 tcttcctcct actcaccatc agcctcctgg ttatggtaca gatacaaact ggactctcag
     1561 gacaaaacga cacctaccct tacgacgtgc ccgactacgc ctcagcatcc agcaacataa
     1621 gcggaggcat tttccttttc ttcgtggcca atgccataat ccacctcttc tgcttcagtt
     1681 gaataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg gtaccgcatc
     1741 ttgctgaaaa actcgagcca tccggaagat ctggcggccg ctctccctat agtgagtcgt
     1801 attacgccgg atggatatgg tgttcaggca caagtgttaa agcagttgat tttattcact
     1861 atgatgaaaa aaacaatgaa tggaacctgc tccaagttaa aaatagagat aataccgaaa
     1921 actcatcgag tagtaagatt agagataata caacaataaa aaaatggttt agaacttact
     1981 cacagcgtga tgctactaat tgggacaatt ttccagatga agtatcatct aagaatttaa
     2041 atgaagaaga cttcagagct tttgttaaaa attatttggc aaaaataata taattcggct
     2101 gcaggggcgg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2161 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2221 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2281 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2341 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2401 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2461 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2521 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2581 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2641 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2701 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2761 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2821 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2881 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2941 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3001 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3061 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3121 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3181 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3241 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4021 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4081 cagtgagcga ggaagcggaa g
//
