LOCUS       sequence-79595-e        7606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRipTAL plasmid for cloning of megaTALs or other TAL-fusions and 
            expression in mammalian cells. RVDs are cloned between Age-Xho sites
            and meganuclease/fusion is cloned between Xba-Sal sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7606)
  TITLE     TALEN Golden Gate vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7606)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCVL RipTAL(Nd154C63) SFFV Age-Xho/Xba-Sal lin...
FEATURES             Location/Qualifiers
     source          1..7606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..235
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             236..416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    463..588
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /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    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2621..3022
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3027..3728
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MASELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             3732..3785
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3792..3818
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3822..3842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(5145..5178)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5383..5400)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5389..5613
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5686..5703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5857..6445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5937..5956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6616..7407,7408..7476))
                     /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     7239..7258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7477..7581)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
       61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
      121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
      181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
      541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
      601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
      661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
      721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
      781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
      841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
      901 aaaagtaaga ccaccgcaca gcaagcggcc ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aatgaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg attacgcgtg tggcatctga agcaccacca gcgagcgaga gctagagaga
     1981 aggaaagcca ccgacttcac cgcctccgag ctgctccggg tcgcgggtct gcagcgtcta
     2041 cggccctccg cgcctacagc tcaagccaca tccgaagggg gagggagccg ggagctgcgc
     2101 gcggggccgc cggggggagg ggtggcaccg cccacgccgg gcggccacga agggcggggc
     2161 agcgggcgcg cgcccggcgg ggggaggggc cgcgcgccgc gcccgctggg aattggggcc
     2221 ctagggggag ggcggaggcg ccgacgaccg cggcacttac cgttcgcggc gtggcgcccg
     2281 gtggtcccca aggggaggga agggggaggc ggggcgagga cagtgaccgg agtctcctca
     2341 gcggtggctt ttctgcttgg cagcctcagc ggctggcgcc aaaaccggac tccgcccact
     2401 tcctcgcccc tgcggtgcga gggtgtggaa tcctccagac gctgggggag ggggagttgg
     2461 gagcttaaaa actagtaccc ctttgggacc actttcagca gcgaactctc ctgtacacca
     2521 ggggtcagtt ccacagacgc gggccagggg tgggtcattg cggcgtgaac aataatttga
     2581 ctagaagttg attcgggtgt ttccggaatt cctagctgca gtaacgccat tttgcaaggc
     2641 atggaaaaat accaaaccaa gaatagagaa gttcagatca agggcgggta catgaaaata
     2701 gctaacgttg ggccaaacag gatatctgcg gtgagcagtt tcggccccgg cccggggcca
     2761 agaacagatg gtcaccgcag tttcggcccc ggcccgaggc caagaacaga tggtccccag
     2821 atatggccca accctcagca gtttcttaag acccatcaga tgtttccagg ctcccccaag
     2881 gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt
     2941 tcgcgcgctt ctgcttcccg agctctataa aagagctcac aacccctcac tcggcgcgcc
     3001 agtcctccga cagactgagt cgcccgatgg ctagcgagct gattaaggag aacatgcaca
     3061 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
     3121 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     3181 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
     3241 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     3301 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     3361 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     3421 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacactg taccccgctg
     3481 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     3541 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     3601 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     3661 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     3721 agcttaattc agagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatccgg
     3781 gcccctgcag gtatccatat gatgtcccag attatgcgcc acctaagaag aaacgcaaag
     3841 tcgtagacct cagaaccctc ggctattcac agcagcagca ggaaaaaatt aagcctaaag
     3901 tgcgaagcac ggttgcccag catcatgagg ccttggtcgg acatggtttt actcacgctg
     3961 atatctgcag aatttctcgg cggagacaga gtttgcgcgt ggtcgctcgg aactaccccg
     4021 agctcgccgc cgccctgcca gaactgactc gcgctcacat agtggatatc gcccggcagc
     4081 ggagcggaga tctggcgctc caagccctgc tgcctgtggc taccgcactt actgccgccc
     4141 cgctgcggct gtccgcctcc cagatcgcca cagtggccca gtacggagag cggccagcaa
     4201 tccaagccct gtataggctg cgaaggaagc ttaccggtgc atgctagctg actgacctcg
     4261 agagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg accaacgacc
     4321 acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg aaaaagggat
     4381 tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa cgcacgtccc
     4441 atcgcgttgc gatatctaga tgacgtagtg atctgatctc agtcgaccga gcatcttacc
     4501 gccatttata cccatatttg ttctgttttt cttgatttgg gtatacattt aaatgttaat
     4561 agaacaaaat ggtggggcaa tcatttacat ttttagggat atgtaattac tagttcaggt
     4621 gtattgccac aagacaaaca tgttaagaaa ctttcccgtt atttacgctc tgttcctgtt
     4681 aatcaacctc tggattacaa aatttgtgaa agattgactg atattcttaa ctatgttgct
     4741 ccttttacgc tgtgtggata tgctgcttta tagcctctgt atctagctat tgcttcccgt
     4801 acggctttcg ttttctcctc cttgtataaa tcctggttgc tgtctctttt agaggagttg
     4861 tggcccgttg tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc aacccccact
     4921 ggctggggca ttgccaccac ctgtcaactc ctttctggga ctttcgcttt ccccctcccg
     4981 atcgccacgg cagaactcat cgccgcctgc cttgcccgct gctggacagg ggctaggttg
     5041 ctgggcactg ataattccgt ggtgttgtca tcgaattcgg taccttttta aaagaaaagg
     5101 ggggactgga agggctaatt cactcccaac gaagacaaga tatcataact tcgtatagca
     5161 tacattatac gaagttataa tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     5221 ccatatgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttgctttttg
     5281 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5341 ggaacccact gcttaagcct caataaagct tgcctcgacc agcctcgact gtgccttcta
     5401 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5461 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5521 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5581 gcaggcatgc tggggatgcg gtgggctcta tggcctgcag ctgcattaat gaatcggcca
     5641 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5701 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5761 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6421 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6541 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6601 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6661 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6721 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6781 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6841 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6901 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6961 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7021 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7081 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7141 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7201 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7261 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7321 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7381 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7441 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7501 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7561 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
