LOCUS       sequence-163607-        6343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4kb HDR template for murine CD90.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  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 6343)
  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: p230_LTJ_4kbCD90.1Template
FEATURES             Location/Qualifiers
     source          1..6343
                     /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(4122..4138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4122..4138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4135..4157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4146..4162
                     /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(4170..4176,4177..4194,4195..4200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4215..4236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4353..4370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4524..5112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4604..4623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5283..6074,6075..6143))
                     /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     5906..5925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6144..6248)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6316..6334
                     /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 caccccatca aaatcctatt
      421 tgagtgtacg gttcggagaa cctcatttat ccggtaaatg tcttttactc tgctctcagg
      481 gagctgaggc aggacatcct gagatacatt gggagaggag atacagtttc aataaaataa
      541 taggttgggt ggaggtacat gcctataatg ccaccactca ggaaatggtg gcagcttcgt
      601 gagtttgagg ccaacccaag aaacatagtg aaaccctgtc agtaaataag taagcaagta
      661 tttgagtatc tactatatgc tagggctgac ctggacatta ggggtcatct tctgaacaaa
      721 ctagtgcttg agggaggtat ttggggtttt tgtttgttta atggatctga atgagttcca
      781 gagactggct acacagcgat atgactgagc ttaacacccc taaagcatac agtcagacca
      841 attagacaat aaaaggtatg tatagcttac caaataaaaa aattgtattt tcaagagagt
      901 gtctgtctgt gtagccctgg ctgttcttga actcactctg tagaccaggc tggcctggaa
      961 atccatctgc ctgcctctgc ctctctgcct ctctgcctct ctgcctctct ctctgcctct
     1021 ctctgcctct ctctgcccct ctctgcccct ctctgcccct ctctgcccct ctctgccgcc
     1081 ctctgccttc tgccctctgc cctctggcct ctggcctctg ccctctgccc tctggcctct
     1141 ggcctctgcc tctgcctctt gagtgctgga atcaaaggtg tgagctctgt aggtcttaag
     1201 ttccagaaga aagtaatgaa gtcacccagc agggaggtgc tcagggacag cacagacaca
     1261 cacccaggac ataggctccc acttccttgg ctttctctga gtggcaaagg accttaggca
     1321 gtgtcactcc ctaagagaag gggataaaga gaggggctga ggtattcatc atgtgctccg
     1381 tggatctcaa gccctcaagg taaatgggga cccacctgtc ctaccagctg gctgacctgt
     1441 agctttcccc accacagaat ccaagtcgga actcttggca ccatgaaccc agccatcagc
     1501 gtcgctctcc tgctctcagg tactgggcaa gggtcagggc tggcattcta aggaatctgg
     1561 cttcctccca tcccgggaag tagcctcttt gccatagtct caggggcaca ggtggttggg
     1621 aggtgcgggg gtggggagtg gggaggagcc tcaacctcac cagtggtggt ctttgacata
     1681 ttagaaactc cataatggat ctaggaactc ctcttgctgg gtggtggtgg ttgtggtaca
     1741 cacctttaat ctcagcactc aggaggcaga gtcaggtgga tctgttagtc tgaagccagc
     1801 ctggtctaca gagcaaattc caggacagcc agagctattc tcaagataga gaatcccttt
     1861 cttgaaaaaa ccatttaaaa acaaaaacaa aagcaacaca ctcctcttga tctcctgttc
     1921 ttgaaacaca ttgttgggac ccagaacttc agtagattga tggaagttgg agtctgcaag
     1981 tggtggaaca tcccaccaat acctcaaggg cgagtgcaaa ccccacatcc ccccagctca
     2041 agtctacttt tcctgcaggt gggaggcccc gggtctgtgt ctccccaaat tcagagaagg
     2101 cactgctgtg ccagtcttgc aggtgtcccg agggcagaag gtgaccagcc tgacagcctg
     2161 cctggtgaac caaaaccttc gcctggactg ccgccatgag aataacacca aggataactc
     2221 catccagcat gagttcagcc tgacccgaga gaagaggaag cacgtgctct caggcaccct
     2281 tgggataccc gagcacacgt accgctcccg cgtcaccctc tccaaccagc cctatatcaa
     2341 ggtccttacc ctagccaact tcaccaccaa ggatgagggc gactactttt gtgagcttcg
     2401 agtctcaggc gcgaatccca tgagctccaa taaaagtatc agtgtgtata gaggtgagac
     2461 tggttcccag aaagataaaa tgtccaggtt agccaggctg gggtagccaa taaaaaaaaa
     2521 aaaaaaaaaa aaaaaaaaaa caggcacctc cattaccctt cccctaactg ctggtctcct
     2581 gggaaactgc tgctgtctat gtgagtgggg caagattagg ggccagaaag ggggagcttg
     2641 tagtaaaagc acagttgagg aaactaaatg ggaaaggcag tacagtggtg attcttgtgg
     2701 tgtggaggtt ctgttacagc atccggtgga gccgctaaga tgagaaagcg ccagctagct
     2761 gccttgaaca gctgacacct gtctttgccc gcctgagtcc tgatctcccc tcctcccggc
     2821 accccttctc tatccacaga caagctggtc aagtgtggcg gcataagcct gctggttcag
     2881 aacacatcct ggatgctgct gctgctgctt tccctctccc tcctccaagc cctggacttc
     2941 atttctctgt gactggttgg gcccaaggag aaacaggagc cctcgaggtc cttcctctgc
     3001 agaggtcttg cttctcccgg tcagctgact ccctccccaa gtccttcaaa tatctcagaa
     3061 catggggaga aacggggacc ttgtccctcc taaggaaccc cagtgctgca tgccatcatc
     3121 ccccccaccc tcgcccccac ccccgccact tctccctcca tgcataccac tagctgtcat
     3181 tttgtactct gtatttattc cagggctgct tctgattatt tagtttgttc tttccctgga
     3241 gacctgttag aacataaggg cgtatggtgg gtaggggagg caggatatca gtccctgggg
     3301 cgagttcctc cctgccaacc aagccagatg cctgaaagag atatggatga gggaagttgg
     3361 actgtgcctg tacctggtac agtcatactc tgttgaaaga atcatcgggg aggggggggg
     3421 gggctcaaga tgggagagct ctgctgagcc tttgtggacc atccaatgag gatgagggct
     3481 tagattctac caggtcattc tcagccacca cacacaagcg ctctgccatc actgaagaag
     3541 ccccctaggg ctcttgggcc agggcacact cagtaaagat gcaggttcag tcagggaatg
     3601 atggggaaag gggtaggagg tgggggaggg atcaccccct cctctaaaac acgagcctgc
     3661 tgtctccaaa ggcctctgcc tgtagtgagg gtggcagaag aagacaagga gccagaactc
     3721 tgactccagg atctaagtcc gtgcaggaag gggatcctag aaccatccgg ttggacccag
     3781 cttaccaagg gagagccttt attcttcttt cccttgcccc tctgtgccag cccctcttgc
     3841 tgtccctgat cccccagaca gcgagagtct tgcaacctgc ctcttccaag acctcctaat
     3901 ctcaggggca ggcggtggag tgagatccgg cgtgcacact ttttggaaga tagctttccc
     3961 aaggatcctc tcccccactg gcagctctgc ctgtcccatc accatgtata ataccaccac
     4021 tgctacagca tctcaccgag gaaagaaaac tgcacaataa aaccaagcct ctggagtgtg
     4081 gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4141 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4201 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4681 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4921 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5161 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5221 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5281 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5341 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5401 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5461 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5521 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5581 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5641 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5701 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5761 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5821 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5881 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5941 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6001 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6061 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6121 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6181 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6241 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6301 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
