LOCUS       sequence-208139-        7493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing cell surface HA-tag and monomeric GFP 
            N-terminaly fused with auxin inducible degron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  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 7493)
  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: pUCHR-CD5HA2-P2A-smAID-mClover
FEATURES             Location/Qualifiers
     source          1..7493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             352..609
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    656..781
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1274..1507
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1692..1736
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2031..2148
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2208..2511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2512..2713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2664..2684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2710..2734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2914..2940
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3028..3084
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(3214..3216,3217..3930)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3256..3277)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3864..3885
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4007..4640
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4689..4705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4689..4705)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4702..4724)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4713..4729
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4737..4743,4744..4761,4762..4767))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4782..4803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4920..4937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      5080..5215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5279..5867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5359..5378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6038..6829,6830..6898))
                     /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     6661..6680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6899..7003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7071..7089
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7127..7149)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7249..7268
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7462..7484
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7476..7493
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7477..7493
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca
       61 gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct
      121 agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt
      181 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt
      241 tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa
      301 ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg
      361 cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg
      421 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
      481 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
      541 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
      601 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
      661 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
      721 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
      781 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
      841 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
      901 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
      961 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     1021 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     1081 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
     1141 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1201 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1261 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1321 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1381 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1441 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1501 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     1561 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     1621 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     1681 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     1741 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     1801 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     1861 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     1921 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     1981 gatctcgacg gtatcggacc ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     2041 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     2101 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     2161 agggacagca gaaatccact ttgggtccga taagcttggg agttccgcgt tacataactt
     2221 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2281 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2341 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2401 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     2461 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2521 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacgggattt ccaagtctcc
     2581 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac ttccaaaatg
     2641 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2701 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     2761 tgacctccat agaagacacc gggaccgatc cagcctccca agcttcgaat tcgcaggtcg
     2821 accaccatga agcgcttcct cttcctccta ctcaccatca gcctcctggt tatggtacag
     2881 atacaaactg gactctcagg acaaaacgac acctaccctt acgacgtgcc cgactacgcc
     2941 tcagcatcca gcaacataag cggaggcatt ttccttttct tcgtggccaa tgccataatc
     3001 cacctcttct gcttcagtgg atccggcgca acaaacttct ctctgctgaa acaagccgga
     3061 gatgtcgaag agaatcctgg accggccaaa cctccggcca aggcacaagt tgtgggatgg
     3121 ccaccggtga gatcataccg gaagaacgtg atggtttcct gccaaaaatc aagcggtggc
     3181 ccggaggcgg cggcgttcgt gaaagtagct agcgtgagca agggcgagga gctgttcacc
     3241 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtc
     3301 cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc
     3361 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcgtggcc
     3421 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3481 gaaggctacg tccaggagcg caccatctct ttcaaggacg acggtaccta caagacccgc
     3541 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3601 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaac
     3661 gtctatatca cggccgacaa gcagaagaac ggcatcaagg ctaacttcaa gatccgccac
     3721 aacgttgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3781 gacggccccg tgctgctgcc cgacaaccac tacctgagcc atcagtccaa gctgagcaaa
     3841 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatt
     3901 acacatggca tggacgagct gtacaagtaa gctcgagtac ctttaagacc aatgacttac
     3961 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     4021 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     4081 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
     4141 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
     4201 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
     4261 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggag
     4321 tacttcaaga actgctgaca tcgagcttgc tacaagggac tttccgctgg ggactttcca
     4381 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatgctg catataagca
     4441 gctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     4501 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4561 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4621 agtgtggaaa atctctagca ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
     4681 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4741 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4801 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4921 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4981 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgcatctc
     5041 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5101 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5161 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagca
     5221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5461 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5701 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7021 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     7081 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     7141 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     7201 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     7261 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7321 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     7381 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     7441 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agt
//
