LOCUS       sequence-204128-        3744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1kb HDR template for murine wildtype Foxp3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3744)
  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 3744)
  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: p249_LTJ_1kbFoxp3wtTemplate
FEATURES             Location/Qualifiers
     source          1..3744
                     /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(1523..1539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1523..1539)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1536..1558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1547..1563
                     /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(1571..1577,1578..1595,1596..1601))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1616..1637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1754..1771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1925..2513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2005..2024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2684..3475,3476..3544))
                     /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     3307..3326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3545..3649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3717..3735
                     /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 acctcgcgaa
      421 tgcatctaga tatccaatgt cgacgaattc acaccaaata tctatggtcc ctggtaaact
      481 cacaacccag gcctgacagg tgggagaggg ttggttagct tggttgagtg cacctgactt
      541 cctggaggtg gggacggaag agagggctct ttggactcac ttggccaggc tcctccctca
      601 tggtgactga atgaagtagc agatgccaaa taggtcaggc ctggctccaa tcccccaaaa
      661 cacggggccc aaggctatgc atttataaca gttgagggct tctaagccta ataaactcag
      721 gtgcagtttc ctcccagctc ttagtggggg tgaggggggt gggggagctc ctttgtaccc
      781 cctggcactg gggtgtggac tccactgacc tgggaaggaa ctattgccat ggcttcccca
      841 gaggtgcctc cgcactgcaa acaggccgcc gtctggagcc tcccgaggag cagaccaggc
      901 cgggagcaca ctgccctgag tactggtggc tacgatgcag caagagctct tgtccattga
      961 cgcctgaggg gttgggaggg gtgtggtaat cgagacactt aagcttgtca tttcattgta
     1021 gcctttcaaa cattagtttg tattttctta ttagttaaac ttgtttttag acaattttgt
     1081 acatgtatgt aatacataca gaccactttc accctatctc ctccctcatc gcttatcttc
     1141 cttcttctca ttcctttccc ccttcatcca tgtctattta ttttggtctt aaaattatta
     1201 ttgccattat tattattatt gacagggtct tactatgtag ccctgactgg cctgcagctc
     1261 tcagagatgc acttcctctg cctcctgagt gtttgcctat tcattttcgt tttgtctgag
     1321 acagtctcac tatatatctc tagttggcct tgcaccttct agcctagttc tacctctgga
     1381 gtgctaggat tataagtgtg tgccaccaca tctggcttcc actttttatt ttatgtatat
     1441 atatatatac ccgggattgg attggatcgg atcccgggcc cgtcgactgc agaggcctgc
     1501 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2341 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2761 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2881 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3121 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3181 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3301 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3361 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3541 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3601 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3661 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3721 ggcgtatcac gaggcccttt cgtc
//
