LOCUS       sequence-163599-        4703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2kb HDR template for murine CD45.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4703)
  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 4703)
  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: p248_LTJ_2kbCD45.1Template
FEATURES             Location/Qualifiers
     source          1..4703
                     /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(2482..2498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2482..2498)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2495..2517)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2506..2522
                     /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(2530..2536,2537..2554,2555..2560))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2575..2596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2713..2730)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2884..3472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2964..2983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3643..4434,4435..4503))
                     /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     4266..4285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4504..4608)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4676..4694
                     /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 tatcggatcc cgggatggtt tacagagcaa gtttcaagac agcccaggat
      481 acacagagaa accctgtctc aaaacaaaca aaacaaaaca aaaagcaaaa aacacaaaac
      541 taaaacaaca taacaacaaa ccatcaaaaa gcagttccta tggattagaa acagcctatt
      601 ttgtattttg tgaaataatt atacataaga ttgctgtttt ttcatttttt tctttcttct
      661 ttttggtatt tctttttaaa ttcaaaatgt tctcattaga gaaaagaatg tagttaaaca
      721 tctaatgttt tctggaaaaa aaaatcaaag ggggaattct agttatgtta ataccaaagt
      781 tgtgatcaca gcacgtggaa ggcaggagta ccttatgtgt tcaaagccag tctatgctgt
      841 atcttgaatt ctgtggcagc cttcacagtg tgagattaca ccttgtttcc aaaaccccaa
      901 acagatcaac aacaagcgtg aaaacagcaa agaaattgtt tttcaaatgg gaaaaatcag
      961 ccgtttatct tgatgtttcc ttgagttgca tttataaaag cagcacacat ttctactgaa
     1021 atctgtggta cgaagaaaat atgttatctt ccctaatgaa tggcaaatat ctgagttgat
     1081 aaacaaactg ccagcaagtg gtgtctacac atatagatat atcaaaacac taataatgtg
     1141 cacatatttg tataattgtg catcagctga aatggagata aattaaaaat taggaatgtg
     1201 actagatcat gaagaagcat cagaccttgg gagagcttag atgtccccta gcgaaatctc
     1261 ctgcttgcaa gactgtactc catgagtctt atttcagagt tgagagggtt cacatctcca
     1321 caccaggaac catcacctaa gagcacctct ccgtttcctc cacagggact gacaagtttt
     1381 cgctacatga ctgcacacca aaagaaaagg ctaatacttc aatttgttta gagtggaaaa
     1441 cagaaaacct tgatttcaga aaatgcaaca gtgacaatat ttcatatgta ctccactgtg
     1501 agccaggtac gatgctgggc agagaagttc tattatcaga aattattcca gacgtggctt
     1561 aaatgttctt tctgtagcct tgtcctcctc acccaccctc agtgatccgc cataaattag
     1621 aataaaataa ccctagtatc tctggcactg aaacaaattc acaacgtaga taaatgaaaa
     1681 gagcaaccca tggatgataa atattatgaa aaattaatta aaagaatttt cttgagtact
     1741 gttatatacc agacattgtt gaaagggtga tgagatgcag gcattataaa agaaagatgg
     1801 tatccatagt ctgtagaaag gaagcatagt attatccaca ccggaggttg ctttctcaga
     1861 ctttcataca tgaaaatcat tggaaagttc aagaatctgc aggtagcaag gaaaacaagt
     1921 gcttctagga gggaacatgg gtttctctag aggttggatc ttttaaaaca tatatttgaa
     1981 ttcagtgagg tggactcttt ctcctctagg catacagcta tgtgccagga cacacttgac
     2041 aagacagagg gaggtacata cacccagagg cggaagagga aatttcactc tgattgtaaa
     2101 cattgtgcta acaggaaatt gctgggctaa gatacaaata ggatgttcac agctggaaca
     2161 aatatatgtt gagttaacat ttttttaaaa aattttaaat aagtcacaat actgatttct
     2221 caacaaaatt gagtatagca atgttatctg caggatcttg gtttattatt tcaccatttt
     2281 cagagacttg accttaacat gaagtcttaa acacatggtg ccataaaaga agcccattta
     2341 ctatgtggca tttcatatac tcatgtgcaa tacttacttc tttcagaaaa taatacaaaa
     2401 tgcattagaa gaaatacatt catacctgaa agatgtcagt tggagctgca gaggcctgca
     2461 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2521 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2581 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2641 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2701 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2761 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2821 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2881 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2941 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3001 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3061 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3121 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3181 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3241 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3301 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3361 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3421 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3481 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3541 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3601 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3661 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3721 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3781 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3841 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3901 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3961 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4021 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4081 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4141 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4201 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4261 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4321 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4381 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4441 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4501 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4561 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4621 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4681 gcgtatcacg aggccctttc gtc
//
