LOCUS       sequence-178613-        3410 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Codon-optimized mKate2 donor for the SapTrap cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3410)
  AUTHORS   Dickinson D, Slabodnick M, Chen A, Goldstein B
  TITLE     SapTrap assembly of repair templates for Cas9-triggered homologous 
            recombination with a self-excising cassette.
  JOURNAL   MicroPubl Biol. 2018 May 1;2018:10.17912/W2KT0N. doi: 
            10.17912/W2KT0N.
  PUBMED    32550377
REFERENCE   2  (bases 1 to 3410)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MicroPubl 
            Biol."; date: "2018-05-1"; volume: "2018:10.17912/W2KT0N. doi"; 
            pages: " 10.17912/W2KT0N"
COMMENT     Sequence Label: pDD375
FEATURES             Location/Qualifiers
     source          1..3410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..36
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     20..36
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     55..75
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        57..75
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     112..128
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     112..128
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    155..179
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1048..1072)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1088..1104)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1089..1105)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1129..1148)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1130..1148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1158..1175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1158..1174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1167..1189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1238..1324
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1416..1443
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1602..1619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1773..2361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1853..1872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2508..3317)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3231..3250
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3309..3328
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agacacgggc cagagctgcc aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
       61 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaact
      121 agtggatccc ccgggctgca ggaattcgat ggggacaagt ttgtacaaaa aagcaggctg
      181 ctcttcgatg gtctccgagc tcattaaaga aaacatgcat atgaagctct acatggaggg
      241 aaccgtcaac aaccaccact tcaagtgcac ctccgaggga gagggaaagc catacgaggg
      301 aacccaaacc atgcgtatca aggccgtcga gggaggacca ctcccattcg ccttcgacat
      361 cctcgccacc tccttcatgt acggatccaa ggtaagttta aacatatata tactaactaa
      421 ccctgattat ttaaattttc agaccttcat caaccacacc caaggaatcc cagacttctt
      481 caagcaatcc ttcccagagg gattcacctg ggagcgtgtc accacctacg aggacggagg
      541 agtcctcacc gccacccaag acacctccct ccaagacgga tgcctcatct acaacgtcaa
      601 ggtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc agatccgtgg
      661 agtcaacttc ccatccaacg gaccagtcat gcaaaagaag accctcggat gggaggcctc
      721 caccgagacc ctctacccag ccgacggagg acttgaggga cgtgccgaca tggccctcaa
      781 gctcgtcgga ggaggacacc tcatctgcaa cctcaaggta agtttaaaca tgattttact
      841 aactaactaa tctgatttaa attttcagac cacctaccgt tccaagaagc cagccaagaa
      901 cctcaagatg ccaggagtct actacgtcga ccgtcgtctt gagcgtatca aggaggccga
      961 caaggagacc tacgtcgagc aacacgaggt cgccgtcgcc cgttactgcg acctcccatc
     1021 caagctcgga caccgtaagc gaagagcacc cagctttctt gtacaaagtg gtccccatca
     1081 agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccaattcgcc ctatagtgag
     1141 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat
     1201 gctagcgtta acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     1261 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     1321 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
     1381 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
     1441 ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc gttgcgctca
     1501 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1561 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1621 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1681 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1741 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1801 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1861 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1921 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1981 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2041 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2101 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2161 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2221 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2281 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2341 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2401 tggaacgacg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     2461 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     2521 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     2581 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     2641 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     2701 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     2761 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     2821 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     2881 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     2941 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3001 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3061 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     3121 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3181 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3241 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc
     3301 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3361 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
//
