LOCUS       sequence-233683-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TAgGFP2 was amplified from pLenti-Origene-Nrf21 and inserted into 
            XhoI and EcoRI digested pLenti-Origene-Nrf21..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  AUTHORS   Lee JY, Chang JK, Dominguez AA, Lee HP, Nam S, Chang J, Varma S, Qi 
            LS, West RB, Chaudhuri O
  TITLE     YAP-independent mechanotransduction drives breast cancer 
            progression.
  JOURNAL   Nat Commun. 2019 Apr 23;10(1):1848. doi: 10.1038/s41467-019-09755-0.
  PUBMED    31015465
REFERENCE   2  (bases 1 to 6970)
  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: "Nat 
            Commun."; date: "2019-04-23"; volume: "10(1):1848. doi"; pages: " 
            10.1038/s41467-019-09755-0"
COMMENT     Sequence Label: pLenti-TagGFP2
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        301..504
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..549
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        530..548
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             563..1276
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     protein_bind    complement(1343..1376)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1432..2020
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1485..1505)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1903..1914)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2023..2039)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2024..2040)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2549..2729
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2791..3379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2871..2890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3456..4115)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4030..4049
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4116..4220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4295..4314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4485..4864
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4866..5064
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5082..5262
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5309..5434
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5931..6164
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6349..6393
                     /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    6691..6808
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        join(6891..6970,1..300)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
       61 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      121 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      181 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      241 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      301 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
      361 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      421 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      481 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc
      541 actatagggc ggccgggaat tcatgagcgg gggcgaggag ctgttcgccg gcatcgtgcc
      601 cgtgctgatc gagctggacg gcgacgtgca cggccacaag ttcagcgtgc gcggcgaggg
      661 cgagggcgac gccgactacg gcaagctgga gatcaagttc atctgcacca ccggcaagct
      721 gcccgtgccc tggcccaccc tggtgaccac cctctgctac ggcatccagt gcttcgcccg
      781 ctaccccgag cacatgaaga tgaacgactt cttcaagagc gccatgcccg agggctacat
      841 ccaggagcgc accatccagt tccaggacga cggcaagtac aagacccgcg gcgaggtgaa
      901 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcaaggact tcaaggagga
      961 cggcaacatc ctgggccaca agctggagta cagcttcaac agccacaacg tgtacatccg
     1021 ccccgacaag gccaacaacg gcctggaggc taacttcaag acccgccaca acatcgaggg
     1081 cggcggcgtg cagctggccg accactacca gaccaacgtg cccctgggcg acggccccgt
     1141 gctgatcccc atcaaccact acctgagcac tcagaccaag atcagcaagg accgcaacga
     1201 ggcccgcgac cacatggtgc tcctggagtc cttcagcgcc tgctgccaca cccacggcat
     1261 ggacgagctg tacaggtccg gactcagata agtttaaacg gccggccgcg gtctgtacaa
     1321 gtaggattcg tcgagggacc taataacttc gtatagcata cattatacga agttatacat
     1381 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct
     1441 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1501 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1561 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1621 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1681 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1741 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1801 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     1861 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     1921 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     1981 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     2041 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     2101 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     2161 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2221 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     2281 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     2341 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     2401 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     2461 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     2521 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     2581 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     2641 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     2701 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
     2761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3421 gaacgaaaac tcacgttaag ggattttggt catgattacg ccccgccctg ccactcatcg
     3481 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     3541 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     3601 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     3661 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     3721 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     3781 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     3841 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaac
     3901 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     3961 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     4021 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     4081 tcaacggtgg tatatccagt gatttttttc tccatactct tcctttttca atattattga
     4141 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4201 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     4261 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4321 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     4381 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     4441 ggcgttttgc gctgcttcgc gatgtacggg ccagatatcg cgttgacatt gattattgac
     4501 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     4561 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     4621 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     4681 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     4741 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     4801 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     4861 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     4921 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     4981 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     5041 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     5101 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5161 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5221 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     5281 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     5341 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     5401 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     5461 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     5521 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     5581 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     5641 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     5701 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     5761 cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat
     5821 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     5881 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     5941 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     6001 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     6061 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     6121 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     6181 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     6241 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     6301 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     6361 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     6421 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     6481 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     6541 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     6601 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     6661 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     6721 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     6781 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     6841 ttggttagta ccgggcccgc tctagacatg tccaatatga ccgccatgtt gacattgatt
     6901 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6961 gttccgcgtt
//
