LOCUS       sequence-219886-        4285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA sequence source for amplifying an HDR template to tag endogenous
            human BATF gene with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4285)
  AUTHORS   Roth TL, Puig-Saus C, Yu R, Shifrut E, Carnevale J, Li PJ, Hiatt J, 
            Saco J, Krystofinski P, Li H, Tobin V, Nguyen DN, Lee MR, Putnam AL,
            Ferris AL, Chen JW, Schickel JN, Pellerin L, Carmody D, 
            Alkorta-Aranburu G, Del Gaudio D, Matsumoto H, Morell M, Mao Y, Cho 
            M, Quadros RM, Gurumurthy CB, Smith B, Haugwitz M, Hughes SH, 
            Weissman JS, Schumann K, Esensten JH, May AP, Ashworth A, Kupfer GM,
            Greeley SAW, Bacchetta R, Meffre E, Roncarolo MG, Romberg N, Herold 
            KC, Ribas A, Leonetti MD, Marson A
  TITLE     Reprogramming human T cell function and specificity with non-viral 
            genome targeting.
  JOURNAL   Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0326-5. doi: 
            10.1038/s41586-018-0326-5.
  PUBMED    29995861
REFERENCE   2  (bases 1 to 4285)
  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: "Nature. 
            2018 Jul 11. pii: 10.1038/s41586-018-0326-5. doi: 
            10.1038/s41586-018-0326-5."
COMMENT     Sequence Label: BATF-GFP HDRT Source (pTR 146)
FEATURES             Location/Qualifiers
     source          1..4285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             881..1393
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVE"
     primer_bind     complement(920..941)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1528..1549
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2064..2080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2064..2080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2077..2099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2088..2104
                     /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(2112..2118,2119..2136,2137..2142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2157..2178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2295..2312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2466..3054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2546..2565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3225..4016,4017..4085))
                     /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     3848..3867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4086..4190)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4258..4276
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggac
      421 tgggctgttt ttctctttta tcttcatcaa ttgaagaggg ctgctgtgtc tttcaaactg
      481 tcggaaggta tctgtatttt tttttctttc acataattct gtaggtcttt tcccagatat
      541 gtatttttct ctctaacttc taactcgctg aagtttccgc ccatgtgact tccagcgtga
      601 gttaccagaa accacaagag agagagagag cgtgcaagcc ccaaagcgag cgacatgtcc
      661 ctttggggag cagtccctct gcaccccaga gtgaggagga cgcaggggtc agaggtggct
      721 acagggcagg cagaggaggc acctgtaggg ggtggtgggc tggtggccca ggagaagtca
      781 ggaagggagc ccagctggtg acaagagagc ccagaggtgc ctggggctga gtgtgagagc
      841 ccggaagatt tcagccatgg gatcgggtgg gactagtggc agcaagggcg aggagctgtt
      901 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      961 cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     1021 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1081 gcagtgcttc agccgctacc ccgaccacat gaagcgccac gacttcttca agtccgccat
     1141 gcccgaaggc tacgtccagg agcgcaccat cagcttcaag gacgacggca cctacaagac
     1201 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1261 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     1321 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1381 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1441 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgtgctgag
     1501 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1561 gatcactgga accggtgctg gaagtggtcc tcacagctcc gacagcagtg actccagctt
     1621 cagccgctct cctccccctg gcaaacaggt agagtcctcc tttttctctc tcctaccttc
     1681 tgattctcct gggggatgga aagagagcca ggcttccttg tcctgcccag ggagctgagg
     1741 atggaggaag tggctcgttg cacgggcact ctgttagact taggacatgg aatttgctac
     1801 taagctgtgc atattggcag agatcctcat ccttccaccc attctgccaa agcccctttt
     1861 tctctccatt ttccaaggct gcctatcacc tctgcctcac tggggttgcc accctaaaaa
     1921 gctttctagg aagcaaagag gaggatgaac atcaaagaat gcagagaaaa gagtctactg
     1981 ttctccaagg ctgtagaaaa gtgagaggag ttttcaagtc ctctagagtc gacctgcagg
     2041 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2101 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2161 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2641 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2881 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4201 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4261 aggcgtatca cgaggccctt tcgtc
//
