LOCUS       sequence-79594-e        7994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRipTAL plasmid compatible with Golden Gate TALEN cloning system. 
            For cloning of megaTALs or other TAL-fusions and expression in 
            mammalian cells. Meganuclease/fusion is cloned between Xba-Sal 
            sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7994)
  TITLE     TALEN Golden Gate vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7994)
  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            "Unpublished"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 Xba-Sal linker
FEATURES             Location/Qualifiers
     source          1..7994
                     /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"
     CDS             complement(3867..4550)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TGKLPSQPIQGLDCWPLSVLGHCGDLGGGQPQRGGSKCGSHRQQGLERQISAPLPGDIH
                     YVSASQFWQGGGELGVVPSDHAQTLSPPRNSADISVSKTMSDQGLMMLGNRASHFRLNF
                     FLLLL"
     primer_bind     4400..4422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4414..4431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4415..4431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4441..4460
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4441..4459
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4507..4527)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4507..4525)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4546..4562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4546..4562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4559..4581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4570..4586
                     /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(4594..4600,4601..4618,4619..4624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4639..4660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(5533..5566)
                     /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(5771..5788)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5777..6001
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6074..6091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6245..6833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6325..6344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7004..7795,7796..7864))
                     /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     7627..7646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7865..7969)
                     /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 ccccctggag acgggcgccg
     4261 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4321 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     4381 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     4441 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cctccagctt
     4501 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4561 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4621 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4681 cgctttccac cggtcgtctc caacgaccac ctcgtcgcct tggcctgcct cggcggacgt
     4741 cctgccatgg atgcagtgaa aaagggattg ccgcacgcgc cggaattgat cagaagagtc
     4801 aatcgccgta ttggcgaacg cacgtcccat cgcgttgcga tatctagatg acgtagtgat
     4861 ctgatctcag tcgaccgagc atcttaccgc catttatacc catatttgtt ctgtttttct
     4921 tgatttgggt atacatttaa atgttaatag aacaaaatgg tggggcaatc atttacattt
     4981 ttagggatat gtaattacta gttcaggtgt attgccacaa gacaaacatg ttaagaaact
     5041 ttcccgttat ttacgctctg ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag
     5101 attgactgat attcttaact atgttgctcc ttttacgctg tgtggatatg ctgctttata
     5161 gcctctgtat ctagctattg cttcccgtac ggctttcgtt ttctcctcct tgtataaatc
     5221 ctggttgctg tctcttttag aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg
     5281 ctctgtgttt gctgacgcaa cccccactgg ctggggcatt gccaccacct gtcaactcct
     5341 ttctgggact ttcgctttcc ccctcccgat cgccacggca gaactcatcg ccgcctgcct
     5401 tgcccgctgc tggacagggg ctaggttgct gggcactgat aattccgtgg tgttgtcatc
     5461 gaattcggta cctttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5521 agacaagata tcataacttc gtatagcata cattatacga agttataatt tatttgtgaa
     5581 atttgtgatg ctattgcttt atttgtaacc atatgtttat ttgtgaaatt tgtgatgcta
     5641 ttgctttatt tgtaaccatt gctttttgct tgtactgggt ctctctggtt agaccagatc
     5701 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5761 cctcgaccag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5821 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5881 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5941 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     6001 gcctgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6061 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6661 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7621 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7981 gaaaagtgcc acct
//
