LOCUS       sequence-204459-        3722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1kb HDR template for murine CD90.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3722)
  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 3722)
  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: p214_LTJ_1kbCD90.1Template
FEATURES             Location/Qualifiers
     source          1..3722
                     /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(1501..1517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1501..1517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1514..1536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1525..1541
                     /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(1549..1555,1556..1573,1574..1579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1594..1615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1732..1749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1903..2491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1983..2002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2662..3453,3454..3522))
                     /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     3285..3304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3523..3627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3695..3713
                     /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 tcgacgggcc cgggatccga tgtcgacatg tgttctgaac cagcaggctt
      481 atgccgccac acttgaccag cttgtctgtg gatagagaag gggtgccggg aggaggggag
      541 atcaggactc aggcgggcaa agacaggtgt cagctgttca aggcagctag ctggcgcttt
      601 ctcatcttag cggctccacc ggatgctgta acagaacctc cacaccacaa gaatcaccac
      661 tgtactgcct ttcccattta gtttcctcaa ctgtgctttt actacaagct ccccctttct
      721 ggcccctaat cttgccccac tcacatagac agcagcagtt tcccaggaga ccagcagtta
      781 ggggaagggt aatggaggtg cctgtttttt tttttttttt tttttttttt tttattggct
      841 accccagcct ggctaacctg gacattttat ctttctggga accagtctca cctctataca
      901 cactgatact tttattggag ctcatgggat tcgcgcctga gactcgaagc tcacaaaagt
      961 agtcgccctc atccttggtg gtgaagttgg ctagggtaag gaccttgata tagggctggt
     1021 tggagagggt gacgcgggag cggtacgtgt gctcgggtat cccaagggtg cctgagagca
     1081 cgtgcttcct cttctctcgg gtcaggctga actcatgctg gatggagtta tccttggtgt
     1141 tattctcatg gcggcagtcc aggcgaaggt tttggttcac caggcaggct gtcaggctgg
     1201 tcaccttctg ccctcgggac acctgcaaga ctggcacagc agtgccttct ctgaatttgg
     1261 ggagacacag acccggggcc tcccacctgc aggaaaagta gacttgagct ggggggatgt
     1321 ggggtttgca ctcgcccttg aggtattggt gggatgttcc accacttgca gactccaact
     1381 tccatcaatc tactgaagtt ctgggtccca acaatgtgaa ttctctagat gcattcgcga
     1441 ggtatcggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg
     1501 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1561 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1621 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1681 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1741 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1801 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2461 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2521 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2581 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2641 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2701 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2761 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2821 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2881 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2941 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3001 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3061 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3121 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3181 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3241 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3301 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3361 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3421 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3481 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3541 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3601 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3661 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3721 tc
//
