LOCUS       sequence-178345-        4098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for Flag 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-CD5Flag2-bglpA
FEATURES             Location/Qualifiers
     source          1..4098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    96..117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(132..137,138..155,156..162)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     enhancer        678..1057
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1058..1261
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1212..1232
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1258..1282
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1561..1584
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1665..1713
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1768..1787)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1769..1787)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2093..2111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2179..2283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2284..2352,2353..3144)
                     /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(2502..2521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3315..3903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3804..3823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4057..4074
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag caattggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      121 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggaat tatgagcgga
      181 taataatttc acacaggagg tttaaacttt aaacatgtca aaagagacgt cttttgttaa
      241 gaatgctgag gaacttgcaa agcaaaaaat ggatgctatt aaccctgaac tttcttcaaa
      301 atttaaattt ttaataaaat tcctgtctca gtttcctgaa gcttgctcta aacctcgttc
      361 aaaaaaaatg cagaataaag ttggtcaaga ggaacatatt gaatatttag ctcgtagttt
      421 tcatgagagt cgattgccaa gaaaacccac gccacctaca acggttcctg atgaggtggt
      481 tagcatagtt cttaatataa gttttaatat acagcctgaa aatcttgaga gaataaaaga
      541 agaacatcga ttttccatgg cagctgagaa tattgtagga gatcttctag aaagatcccc
      601 gcggccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
      661 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      721 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      781 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      841 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      901 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      961 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1021 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     1081 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     1141 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     1201 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1261 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     1321 aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc
     1381 ccgtgccaag agtgacgtaa gtaccgccta tagagtctat aggcccaccc ccttggcttc
     1441 ttatgcgacg gatctgcctg caggtcgacc accatgaagc gcttcctctt cctcctactc
     1501 accatcagcc tcctggttat ggtacagata caaactggac tctcaggaca aaacgacacc
     1561 gactacaaag acgatgacga caagtcagca tccagcaaca taagcggagg cattttcctt
     1621 ttcttcgtgg ccaatgccat aatccacctc ttctgcttca gttgaataaa atatctttat
     1681 tttcattaca tctgtgtgtt ggttttttgt gtggtaccgc atcttgctga aaaactcgag
     1741 ccatccggaa gatctggcgg ccgctctccc tatagtgagt cgtattacgc cggatggata
     1801 tggtgttcag gcacaagtgt taaagcagtt gattttattc actatgatga aaaaaacaat
     1861 gaatggaacc tgctccaagt taaaaataga gataataccg aaaactcatc gagtagtaag
     1921 attagagata atacaacaat aaaaaaatgg tttagaactt actcacagcg tgatgctact
     1981 aattgggaca attttccaga tgaagtatca tctaagaatt taaatgaaga agacttcaga
     2041 gcttttgtta aaaattattt ggcaaaaata atataattcg gctgcagggg cggcctcgtg
     2101 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2161 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2221 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2281 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2341 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2401 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2461 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2521 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2581 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2641 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2701 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2761 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2821 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2881 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2941 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3001 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3061 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3121 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3181 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3421 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3961 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4021 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4081 gaagagcgcc caatacgc
//
