LOCUS       sequence-163600-        6703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4kb HDR template for murine CD45.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  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 6703)
  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: p243_LTJ_4kbCD45.1Template
FEATURES             Location/Qualifiers
     source          1..6703
                     /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(4482..4498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4482..4498)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4495..4517)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4506..4522
                     /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(4530..4536,4537..4554,4555..4560))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4575..4596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4713..4730)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4884..5472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4964..4983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5643..6434,6435..6503))
                     /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     6266..6285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6504..6608)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6676..6694
                     /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 cgggttaatt caccaagtat tattgaattg ttgataacag
      481 acagatatat cataaacccg agttaatgat aatgaacaaa atagactagt tatggttctc
      541 atggactttt caatttagtt ggaaagatag ataccaaacc aataattatc atagtataaa
      601 ctgggatctt ttataggaga aaggatttca aaataatgat gtaggggtac atgatacagt
      661 tcttctattt ttcatattcc tggagatact gtgctacaac ctacaatctg ctttggtgat
      721 attttggtat attacctctg ctatatgcta attattcttt tacttctgct tgctgagtac
      781 tgaatttgaa acccaaaata cacttatttc atcagatcat tctctgtaca aaagaatatg
      841 ttaagacatt tgtatccaat gtagtatcat tcattctcga tgatgattgt actaatttcc
      901 tttttttttt tttccagctg ccatgtttgg gaacattact gtgaattaca cctatgaatc
      961 tagtaatcag acttttaagg cagacctcaa agatgtccaa aatgctaagt gtggaaatga
     1021 ggattgtgaa aacgtgttaa ataatctaga agaatgctca cagataaaaa acatcagtgt
     1081 gtctaatgac tcatgtgctc cagctacaac tatagattta tatgtaccac caggtgaatg
     1141 tcaatttctc tctatctggg tatttagagc aacagaagga tttcatgtat gtgttcctgt
     1201 gcttcttcaa gctaatgtgg tcagaaaaaa gaaccgaggg aaagcaagtg agctttgata
     1261 agagtgtgtt tcctgctgag aaccccggag agcagagaga aatacacact tattccttag
     1321 gaccaccacc tggctctctt atttctttta taaagagatt cccagctggg aagtggtgtc
     1381 cctcaccttc aatctcagca cttcagaggc agaggcaggc agatctcagg gttccaaggc
     1441 agccatggtt tacagagcaa gtttcaagac agcccaggat acacagagaa accctgtctc
     1501 aaaacaaaca aaacaaaaca aaaagcaaaa aacacaaaac taaaacaaca taacaacaaa
     1561 ccatcaaaaa gcagttccta tggattagaa acagcctatt ttgtattttg tgaaataatt
     1621 atacataaga ttgctgtttt ttcatttttt tctttcttct ttttggtatt tctttttaaa
     1681 ttcaaaatgt tctcattaga gaaaagaatg tagttaaaca tctaatgttt tctggaaaaa
     1741 aaaatcaaag ggggaattct agttatgtta ataccaaagt tgtgatcaca gcacgtggaa
     1801 ggcaggagta ccttatgtgt tcaaagccag tctatgctgt atcttgaatt ctgtggcagc
     1861 cttcacagtg tgagattaca ccttgtttcc aaaaccccaa acagatcaac aacaagcgtg
     1921 aaaacagcaa agaaattgtt tttcaaatgg gaaaaatcag ccgtttatct tgatgtttcc
     1981 ttgagttgca tttataaaag cagcacacat ttctactgaa atctgtggta cgaagaaaat
     2041 atgttatctt ccctaatgaa tggcaaatat ctgagttgat aaacaaactg ccagcaagtg
     2101 gtgtctacac atatagatat atcaaaacac taataatgtg cacatatttg tataattgtg
     2161 catcagctga aatggagata aattaaaaat taggaatgtg actagatcat gaagaagcat
     2221 cagaccttgg gagagcttag atgtccccta gcgaaatctc ctgcttgcaa gactgtactc
     2281 catgagtctt atttcagagt tgagagggtt cacatctcca caccaggaac catcacctaa
     2341 gagcacctct ccgtttcctc cacagggact gacaagtttt cgctacatga ctgcacacca
     2401 aaagaaaagg ctaatacttc aatttgttta gagtggaaaa cagaaaacct tgatttcaga
     2461 aaatgcaaca gtgacaatat ttcatatgta ctccactgtg agccaggtac gatgctgggc
     2521 agagaagttc tattatcaga aattattcca gacgtggctt aaatgttctt tctgtagcct
     2581 tgtcctcctc acccaccctc agtgatccgc cataaattag aataaaataa ccctagtatc
     2641 tctggcactg aaacaaattc acaacgtaga taaatgaaaa gagcaaccca tggatgataa
     2701 atattatgaa aaattaatta aaagaatttt cttgagtact gttatatacc agacattgtt
     2761 gaaagggtga tgagatgcag gcattataaa agaaagatgg tatccatagt ctgtagaaag
     2821 gaagcatagt attatccaca ccggaggttg ctttctcaga ctttcataca tgaaaatcat
     2881 tggaaagttc aagaatctgc aggtagcaag gaaaacaagt gcttctagga gggaacatgg
     2941 gtttctctag aggttggatc ttttaaaaca tatatttgaa ttcagtgagg tggactcttt
     3001 ctcctctagg catacagcta tgtgccagga cacacttgac aagacagagg gaggtacata
     3061 cacccagagg cggaagagga aatttcactc tgattgtaaa cattgtgcta acaggaaatt
     3121 gctgggctaa gatacaaata ggatgttcac agctggaaca aatatatgtt gagttaacat
     3181 ttttttaaaa aattttaaat aagtcacaat actgatttct caacaaaatt gagtatagca
     3241 atgttatctg caggatcttg gtttattatt tcaccatttt cagagacttg accttaacat
     3301 gaagtcttaa acacatggtg ccataaaaga agcccattta ctatgtggca tttcatatac
     3361 tcatgtgcaa tacttacttc tttcagaaaa taatacaaaa tgcattagaa gaaatacatt
     3421 catacctgaa agatgtcagt tggacaacct tcgtgcccaa acaaattaca catgtgtagc
     3481 agaaatctta tatcgcggtg taaaactcgt caaaaatgtt ataaatgtgc agacagattt
     3541 ggggagtaag tatatcgttt atgtttataa aataaataaa atttcttttt cttttatggg
     3601 tttactttta tttttggaga gtcatagaaa ataatggtat agaattacag aaagcatatc
     3661 agtgaagaaa aaagtttaaa tgcactttaa ggtcaatttt aaaaactagg ttatacaaac
     3721 ctaactaaag ttctttttct gttgttcaaa ttgtgtctgc cgtaagtctg gtagaaacac
     3781 ttgaaatggt ttacatagat tattcccatt ttattcattt tctcctccct tctttttata
     3841 acaccatttg ttctgtattc taacacttct ttcacaatta tgctctgctt ctaatagaaa
     3901 taaaataagc aaagaaacta ataaaactca agcaaatcta catgtttaaa cataaaatac
     3961 ggagtatagt tatcaaattt tggcaataga attaataacc ggataaacct tattaaatag
     4021 aaaggggttc tacttgggaa gtcagatgtg aaattttcaa attaaatgta acatttgatg
     4081 gtaaatgcta attttttaaa atggtgatta atgaataatc aaccataaag aagacaaaag
     4141 atgtcattgc tcacaagcag taaagacatt tttagaagga agagaagaga agcagtagga
     4201 aaggaggaag ggagagatag atgatagata gataatagat agatagatag atagatagat
     4261 agatagatag agacagacag acagacagaa agaaagagag agacagaaac agagagacac
     4321 aaaaaaagag gaaagagaag actgggagaa agggaaagta agggatggag aaaaaaggga
     4381 gagagataaa aagaagacaa aaacataaat gaaagcaagg aatgatgata tatatctgtt
     4441 atgtgctgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4501 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4561 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4621 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4681 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4741 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4801 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4861 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4921 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4981 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5041 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5101 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5161 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5221 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5281 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5341 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5401 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5461 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5521 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5581 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5641 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5701 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5761 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5821 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5881 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5941 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6001 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6061 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6121 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6181 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6241 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6301 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6361 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6421 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6481 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6541 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6601 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6661 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
