LOCUS       sequence-163602-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2kb HDR template for murine wildtype Foxp3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4675)
  AUTHORS   Kornete M, Marone R, Jeker LT
  TITLE     Highly Efficient and Versatile Plasmid-Based Gene Editing in Primary
            T Cells.
  JOURNAL   J Immunol. 2018 Feb 14. pii: jimmunol.1701121. doi: 
            10.4049/jimmunol.1701121.
  PUBMED    29445007
REFERENCE   2  (bases 1 to 4675)
  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: "J Immunol. 
            2018 Feb 14. pii: jimmunol.1701121. doi: 10.4049/jimmunol.1701121."
COMMENT     Sequence Label: p250_LTJ_2kbFoxp3wtTemplate
FEATURES             Location/Qualifiers
     source          1..4675
                     /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"
     primer_bind     complement(2454..2470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2454..2470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2467..2489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2478..2494
                     /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(2502..2508,2509..2526,2527..2532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2547..2568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2685..2702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2856..3444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2936..2955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3615..4406,4407..4475))
                     /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     4238..4257
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4476..4580)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4648..4666
                     /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 ccccgggact ccagttccag
      421 tggatccaat gcacttttct gccttccaca ggtaccaggc acacatgcga tgcccagaca
      481 tgcatgcagg caaaactccc gtatacctaa aataaaatgc aagctgactt ggcagtaatc
      541 tcagcccatc ctgtgctaca tagtacatgt tagactagcc tgtactacat gctacatagt
      601 acatgttaga ctagcctgta ctacatgcta catagtacat gttagactag cctgtactac
      661 agagcaagag cccacctaca taaatatcca accaagcaag caatcatttt ttaaagtaaa
      721 atggaagact cagtgtggtg gcgcacgcac gcctttaatc ctagaactcg ggaggcagat
      781 gcaggcagat ctctgtgagt tcgaagccag tctggtctac agagcctggt ctatacactg
      841 agctccagga cagccaagac tacacagaga aaccctgtct ggaagaaaaa aaaaatatat
      901 atatatatat atatatatat acataaaata aaaagtggaa gccagatgtg gtggcacaca
      961 cttataatcc tagcactcca gaggtagaac taggctagaa ggtgcaaggc caactagaga
     1021 tatatagtga gactgtctca gacaaaacga aaatgaatag gcaaacactc aggaggcaga
     1081 ggaagtgcat ctctgagagc tgcaggccag tcagggctac atagtaagac cctgtcaata
     1141 ataataataa tggcaataat aattttaaga ccaaaataaa tagacatgga tgaaggggga
     1201 aaggaatgag aagaaggaag ataagcgatg agggaggaga tagggtgaaa gtggtctgta
     1261 tgtattacat acatgtacaa aattgtctaa aaacaagttt aactaataag aaaatacaaa
     1321 ctaatgtttg aaaggctaca atgaaatgac aagcttaagt gtctcgatta ccacacccct
     1381 cccaacccct caggcgtcaa tggacaagag ctcttgctgc atcgtagcca ccagtactca
     1441 gggcagtgtg ctcccggcct ggtctgctcc tcgggaggct ccagacggcg gcctgtttgc
     1501 agtgcggagg cacctctggg gaagccatgg caatagttcc ttcccaggtc agtggagtcc
     1561 acaccccagt gccagggggt acaaaggagc tcccccaccc ccctcacccc cactaagagc
     1621 tgggaggaaa ctgcacctga gtttattagg cttagaagcc ctcaactgtt ataaatgcat
     1681 agccttgggc cccgtgtttt gggggattgg agccaggcct gacctatttg gcatctgcta
     1741 cttcattcag tcaccatgag ggaggagcct ggccaagtga gtccaaagag ccctctcttc
     1801 cgtccccacc tccaggaagt caggtgcact caaccaagct aaccaaccct ctcccacctg
     1861 tcaggcctgg gttgtgagtt taccagggac catagatatt tggtgtcagg ctggctatgc
     1921 cacttgagct gcttacatgc ctttgatgta caaattactt gactcctttt taaagtgagg
     1981 agagctattt ggcaggagta ctgcaaagaa gacacagctt acggcgggta ctcagtaaac
     2041 agtactatgt gtgagcatag actgtccctc cccccttggt gctagtggta ggaattgaga
     2101 ccttggattc ctgatgcaga caaaggtggg gtagggggtg aggaggccaa aggctctgat
     2161 ctatgccaac cttctgcaga gttcttccac aacatggact acttcaagta ccacaatatg
     2221 cgaccccctt tcacctatgc cacccttatc cgatgggtaa gcagggcaat agaggcccag
     2281 cagctggtgg gcggcagggg gggagttgtg gtggggagtg cttgcctcct acattgcacc
     2341 aagagcagaa ttcacccatt aacaaacctc agctctgagg agccccaaga tgtgatcctt
     2401 cttgatgaat tcgtcgactg cagaggcctg catgcaagct tggcgtaatc atggtcatag
     2461 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2521 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2581 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2641 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2701 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2761 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2821 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2881 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2941 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3001 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3061 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3121 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3181 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3241 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3301 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3361 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3421 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3481 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3541 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3601 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3661 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3721 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3781 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3841 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3901 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3961 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4021 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4081 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4141 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4201 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4261 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4321 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4381 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4441 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4501 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4561 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4621 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
