LOCUS       sequence-75354-e        8398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTAL plasmid compatible with the Golden Gate TALEN cloning system, 
            for cloning of TALENs and expression in mammalian cells. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8398)
  TITLE     TALEN Golden Gate vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8398)
  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            "Unpublished"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: pTALEN(Nd154C63) Ef1a
FEATURES             Location/Qualifiers
     source          1..8398
                     /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        2632..2843
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2852..2861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2858..3559
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MASELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             3563..3616
                     /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             3623..3649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3653..3673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3974..4381)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     4231..4253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4245..4262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4246..4262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4272..4291
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4272..4290
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4338..4358)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4338..4356)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4377..4393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4377..4393)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4390..4412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4401..4417
                     /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(4425..4431,4432..4449,4450..4455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4470..4491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             4683..5270
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     protein_bind    complement(5937..5970)
                     /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(6175..6192)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6181..6405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6478..6495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6649..7237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6729..6748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7408..8199,8200..8268))
                     /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     8031..8050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8269..8373)
                     /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 caggctccgg tgcccgtcag tgggcagagc
     2641 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     2701 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     2761 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     2821 acgggtttgc cgccagaaca caggatctgc cgccaccatg gctagcgagc tgattaagga
     2881 gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac
     2941 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     3001 gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa
     3061 gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg
     3121 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     3181 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac
     3241 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacact
     3301 gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg
     3361 gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct
     3421 caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa
     3481 cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa
     3541 actggggcac aagcttaatt cagagggcag aggaagtctt ctaacatgcg gtgacgtgga
     3601 ggagaatccg ggcccctgca ggtatccata tgatgtccca gattatgcgc cacctaagaa
     3661 gaaacgcaaa gtcgtggatc tacgcacgct cggctacagt cagcagcagc aagagaagat
     3721 caaaccgaag gtgcgttcga cagtggcgca gcaccacgag gcactggtgg gccatgggtt
     3781 tacacacgcg cacatcgttg cgctcagcca acacccggca gcgttaggga ccgtcgctgt
     3841 cacgtatcag cacataatca cggcgttgcc agaggcgaca cacgaagaca tcgttggcgt
     3901 cggcaaacag tggtccggcg cacgcgccct ggaggccttg ctcacggatg cgggggagtt
     3961 gagaggtccg ccgttacagt tggacacagg ccaacttgtg aagattgcaa aacgtggcgg
     4021 cgtgaccgca atggaggcag tgcatgcatc gcgcaatgca ctgacgggtg cccccctgga
     4081 gacgggcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     4141 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4201 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4261 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg
     4321 tcctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     4381 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4441 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4501 cgctcactgc ccgctttcca ccggtcgtct ccaacgacca cctcgtcgcc ttggcctgcc
     4561 tcggcggacg tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga
     4621 tcagaagagt caatcgccgt attggcgaac gcacgtccca tcgcgttgcg atatctagat
     4681 cccagctagt gaaatctgaa ttggaagaga agaaatctga acttagacat aaattgaaat
     4741 atgtgccaca tgaatatatt gaattgattg aaatcgcaag aaattcaact caggatagaa
     4801 tccttgaaat gaaggtgatg gagttcttta tgaaggttta tggttatcgt ggtaaacatt
     4861 tgggtggatc aaggaaacca gacggagcaa tttatactgt cggatctcct attgattacg
     4921 gtgtgatcgt tgatactaag gcatattcag gaggttataa tcttccaatt ggtcaagcag
     4981 atgaaatgca aagatatgtc gaagagaatc aaacaagaaa caagcatatc aaccctaatg
     5041 aatggtggaa agtctatcca tcttcagtaa cagaatttaa gttcttgttt gtgagtggtc
     5101 atttcaaagg aaactacaaa gctcagctta caagattgaa tcatatcact aattgtaatg
     5161 gagctgttct tagtgtagaa gagcttttga ttggtggaga aatgattaaa gctggtacat
     5221 tgacacttga ggaagtgaga aggaaattta ataacggtga gataaacttt taagtcgacc
     5281 gagcatctta ccgccattta tacccatatt tgttctgttt ttcttgattt gggtatacat
     5341 ttaaatgtta atagaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt
     5401 actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc
     5461 tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt
     5521 aactatgttg ctccttttac gctgtgtgga tatgctgctt tatagcctct gtatctagct
     5581 attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt
     5641 ttagaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac
     5701 gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct
     5761 ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca
     5821 ggggctaggt tgctgggcac tgataattcc gtggtgttgt catcgaattc ggtacctttt
     5881 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatcataa
     5941 cttcgtatag catacattat acgaagttat aatttatttg tgaaatttgt gatgctattg
     6001 ctttatttgt aaccatatgt ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6061 cattgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6121 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgcctcga ccagcctcga
     6181 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6241 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6301 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6361 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcctgc agctgcatta
     6421 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6481 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6541 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6601 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6661 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6721 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6781 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6841 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6901 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6961 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7021 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7081 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7141 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7201 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7261 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7321 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7381 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7441 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7501 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7561 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7621 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7681 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7741 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7801 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7861 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7921 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7981 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8041 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8101 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8161 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8221 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8341 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacct
//
