LOCUS       sequence-163606-        4664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2kb HDR template for murine CD90.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4664)
  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 4664)
  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: p229_LTJ_2kbCD90.1Template
FEATURES             Location/Qualifiers
     source          1..4664
                     /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(2443..2459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2443..2459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2456..2478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2467..2483
                     /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(2491..2497,2498..2515,2516..2521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2536..2557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2674..2691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2845..3433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2925..2944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3604..4395,4396..4464))
                     /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     4227..4246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4465..4569)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4637..4655
                     /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 caaatgggga cccacctgtc
      421 ctaccagctg gctgacctgt agctttcccc accacagaat ccaagtcgga actcttggca
      481 ccatgaaccc agccatcagc gtcgctctcc tgctctcagg tactgggcaa gggtcagggc
      541 tggcattcta aggaatctgg cttcctccca tcccgggaag tagcctcttt gccatagtct
      601 caggggcaca ggtggttggg aggtgcgggg gtggggagtg gggaggagcc tcaacctcac
      661 cagtggtggt ctttgacata ttagaaactc cataatggat ctaggaactc ctcttgctgg
      721 gtggtggtgg ttgtggtaca cacctttaat ctcagcactc aggaggcaga gtcaggtgga
      781 tctgttagtc tgaagccagc ctggtctaca gagcaaattc caggacagcc agagctattc
      841 tcaagataga gaatcccttt cttgaaaaaa ccatttaaaa acaaaaacaa aagcaacaca
      901 ctcctcttga tctcctgttc ttgaaacaca ttgttgggac ccagaacttc agtagattga
      961 tggaagttgg agtctgcaag tggtggaaca tcccaccaat acctcaaggg cgagtgcaaa
     1021 ccccacatcc ccccagctca agtctacttt tcctgcaggt gggaggcccc gggtctgtgt
     1081 ctccccaaat tcagagaagg cactgctgtg ccagtcttgc aggtgtcccg agggcagaag
     1141 gtgaccagcc tgacagcctg cctggtgaac caaaaccttc gcctggactg ccgccatgag
     1201 aataacacca aggataactc catccagcat gagttcagcc tgacccgaga gaagaggaag
     1261 cacgtgctct caggcaccct tgggataccc gagcacacgt accgctcccg cgtcaccctc
     1321 tccaaccagc cctatatcaa ggtccttacc ctagccaact tcaccaccaa ggatgagggc
     1381 gactactttt gtgagcttcg agtctcaggc gcgaatccca tgagctccaa taaaagtatc
     1441 agtgtgtata gaggtgagac tggttcccag aaagataaaa tgtccaggtt agccaggctg
     1501 gggtagccaa taaaaaaaaa aaaaaaaaaa aaaaaaaaaa caggcacctc cattaccctt
     1561 cccctaactg ctggtctcct gggaaactgc tgctgtctat gtgagtgggg caagattagg
     1621 ggccagaaag ggggagcttg tagtaaaagc acagttgagg aaactaaatg ggaaaggcag
     1681 tacagtggtg attcttgtgg tgtggaggtt ctgttacagc atccggtgga gccgctaaga
     1741 tgagaaagcg ccagctagct gccttgaaca gctgacacct gtctttgccc gcctgagtcc
     1801 tgatctcccc tcctcccggc accccttctc tatccacaga caagctggtc aagtgtggcg
     1861 gcataagcct gctggttcag aacacatcct ggatgctgct gctgctgctt tccctctccc
     1921 tcctccaagc cctggacttc atttctctgt gactggttgg gcccaaggag aaacaggagc
     1981 cctcgaggtc cttcctctgc agaggtcttg cttctcccgg tcagctgact ccctccccaa
     2041 gtccttcaaa tatctcagaa catggggaga aacggggacc ttgtccctcc taaggaaccc
     2101 cagtgctgca tgccatcatc ccccccaccc tcgcccccac ccccgccact tctccctcca
     2161 tgcataccac tagctgtcat tttgtactct gtatttattc cagggctgct tctgattatt
     2221 tagtttgttc tttccctgga gacctgttag aacataaggg cgtatggtgg gtaggggagg
     2281 caggatatca gtccctgggg cgagttcctc cctgccaacc aagccagatg cctgaaagag
     2341 atatggatga gggaagttgg actgtgcctg tacctggtac agtcatactc tgttgaaaga
     2401 agtcgactgc agaggcctgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     2461 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2581 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2641 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2701 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2761 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2821 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2881 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2941 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3001 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3061 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3121 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3181 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3241 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3301 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3361 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3421 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3481 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3541 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3601 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3661 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3721 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3781 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3841 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3901 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3961 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4021 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4081 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4141 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4201 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4261 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4321 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4381 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4441 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4501 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4561 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4621 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
