LOCUS       sequence-75688-e        6862 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pTAL 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 6862)
  TITLE     TALEN Golden Gate vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6862)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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 TAL(Nd154C63) T7 Age-Xho/Xba-Sal linkers
FEATURES             Location/Qualifiers
     source          1..6862
                     /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"
     primer_bind     3001..3020
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3001..3019
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3024..3033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3045..3071
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3075..3095
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4401..4434)
                     /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(4639..4656)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4645..4869
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4942..4959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5113..5701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5193..5212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5872..6663,6664..6732))
                     /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     6495..6514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6733..6837)
                     /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 taatacgact cactataggg gccgccacca tgggatcctg caggtatcca tatgatgtcc
     3061 cagattatgc gccacctaag aagaaacgca aagtcgtgga tctacgcacg ctcggctaca
     3121 gtcagcagca gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg
     3181 aggcactggt gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg
     3241 cagcgttagg gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga
     3301 cacacgaaga catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct
     3361 tgctcacgga tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg
     3421 tgaagattgc aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg
     3481 cactgaccgg tgcatgctag ctgactgacc tcgagagcat tgtggcccag ctgagccggc
     3541 ctgatccggc gttggccgcg ttgaccaacg accacctcgt cgccttggcc tgcctcggcg
     3601 gacgtcctgc catggatgca gtgaaaaagg gattgccgca cgcgccggaa ttgatcagaa
     3661 gagtcaatcg ccgtattggc gaacgcacgt cccatcgcgt tgcgatatct agatgagacg
     3721 tagtgatctg acgtctcgtc gaccgagcat cttaccgcca tttataccca tatttgttct
     3781 gtttttcttg atttgggtat acatttaaat gttaatagaa caaaatggtg gggcaatcat
     3841 ttacattttt agggatatgt aattactagt tcaggtgtat tgccacaaga caaacatgtt
     3901 aagaaacttt cccgttattt acgctctgtt cctgttaatc aacctctgga ttacaaaatt
     3961 tgtgaaagat tgactgatat tcttaactat gttgctcctt ttacgctgtg tggatatgct
     4021 gctttatagc ctctgtatct agctattgct tcccgtacgg ctttcgtttt ctcctccttg
     4081 tataaatcct ggttgctgtc tcttttagag gagttgtggc ccgttgtccg tcaacgtggc
     4141 gtggtgtgct ctgtgtttgc tgacgcaacc cccactggct ggggcattgc caccacctgt
     4201 caactccttt ctgggacttt cgctttcccc ctcccgatcg ccacggcaga actcatcgcc
     4261 gcctgccttg cccgctgctg gacaggggct aggttgctgg gcactgataa ttccgtggtg
     4321 ttgtcatcga attcggtacc tttttaaaag aaaagggggg actggaaggg ctaattcact
     4381 cccaacgaag acaagatatc ataacttcgt atagcataca ttatacgaag ttataattta
     4441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat atgtttattt gtgaaatttg
     4501 tgatgctatt gctttatttg taaccattgc tttttgcttg tactgggtct ctctggttag
     4561 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     4621 aaagcttgcc tcgaccagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4681 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4741 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4801 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4861 gctctatggc ctgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4921 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5281 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5521 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5821 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5881 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5941 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6001 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6061 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6841 atttccccga aaagtgccac ct
//
