LOCUS       sequence-214600-        7552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing cell surface c-Myc-tag and monomeric 
            GFP C-terminaly fused with auxin inducible degron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7552)
  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 7552)
  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-mClover-smAID-P2A-CD5cmyc2-bglpA
FEATURES             Location/Qualifiers
     source          1..7552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             350..607
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    654..779
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1272..1505
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1690..1734
                     /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    2029..2146
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2206..2509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2510..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             join(2819..2821,2822..2824,2825..3535)
                     /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="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2864..2885)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3472..3493
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3671..3727
                     /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             3812..3841
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3922..3970
                     /note="synthetic polyadenylation signal"
     LTR             4067..4700
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4749..4765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4749..4765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4762..4784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4773..4789
                     /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(4797..4803,4804..4821,4822..4827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4842..4863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4980..4997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      5140..5275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5338..5926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5418..5437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6097..6888,6889..6957))
                     /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     6720..6739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6958..7062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7130..7148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7186..7208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7308..7327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7521..7543
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7535..7552
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7536..7552
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattgatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
       61 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
      121 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
      181 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
      241 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
      301 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
      361 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
      421 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      481 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
      541 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      601 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
      661 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
      721 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
      781 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
      841 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
      901 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
      961 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1021 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1081 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     1141 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1201 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1261 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1321 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1381 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1441 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1501 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     1561 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     1621 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     1681 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     1741 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     1801 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     1861 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     1921 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     1981 tctcgacggt atcggaccct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2041 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2101 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2161 ggacagcaga aatccacttt gggtccgata agcttgggag ttccgcgtta cataacttac
     2221 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2281 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2341 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2401 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2461 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2521 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2581 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2641 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2701 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     2761 tgacctccat agaagacacc gggaccgatc cagcctccga attcgcaggt cgaccaccat
     2821 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2881 cgacgtaaac ggccacaagt tcagcgtccg cggcgagggc gagggcgatg ccaccaacgg
     2941 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3001 cgtgaccacc ttcggctacg gcgtggcctg cttcagccgc taccccgacc acatgaagca
     3061 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatctcttt
     3121 caaggacgac ggtacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3181 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3241 gctggagtac aacttcaaca gccacaacgt ctatatcacg gccgacaagc agaagaacgg
     3301 catcaaggct aacttcaaga tccgccacaa cgttgaggac ggcagcgtgc agctcgccga
     3361 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3421 cctgagccat cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3481 gctggagttc gtgaccgccg ccgggattac acatggcatg gacgagctgt acaagccagc
     3541 caaacctccg gccaaggcac aagttgtggg atggccaccg gtgagatcat accggaagaa
     3601 cgtgatggtt tcctgccaaa aatcaagcgg tggcccggag gcggcggcgt tcgtgaaagt
     3661 aggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc
     3721 tggaccgaag cgcttcctct tcctcctact caccatcagc ctcctggtta tggtacagat
     3781 acaaactgga ctctcaggac aaaacgacac cgagcagaag ctgatcagcg aggaagacct
     3841 gtcagcatcc agcaacataa gcggaggcat tttccttttc ttcgtggcca atgccataat
     3901 ccacctcttc tgcttcagtt gaataaaata tctttatttt cattacatct gtgtgttggt
     3961 tttttgtgtg gtaccgcatc ttgctgaaaa actcgagtac ctttaagacc aatgacttac
     4021 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     4081 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     4141 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
     4201 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
     4261 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
     4321 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggag
     4381 tacttcaaga actgctgaca tcgagcttgc tacaagggac tttccgctgg ggactttcca
     4441 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatgctg catataagca
     4501 gctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     4561 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4621 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4681 agtgtggaaa atctctagca ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
     4741 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4801 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgcatctc
     5101 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5161 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5221 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagat
     5281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6241 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7081 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7141 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7201 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7261 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7321 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7381 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7441 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7501 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gt
//
