LOCUS       sequence-439410-        2338 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Backbone with MCS flanked by attP and Lox sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2338)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 2338)
  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"
COMMENT     Sequence Label: pSwitch-MCS-AttP-2Lox
FEATURES             Location/Qualifiers
     source          1..2338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..21)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5..21)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(123..142)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             134..943
                     /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     complement(201..220)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1090..1678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1579..1598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1832..1849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(1875..1922)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(1938..1971)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1983..2052
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     misc_feature    2068..2175
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2092..2108
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2092..2108
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2142..2158)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2143..2159)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(2277..2310)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2319..2338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2320..2338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
       61 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
      121 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
      181 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
      241 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
      301 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
      361 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
      421 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
      481 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
      541 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
      601 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
      661 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
      721 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
      781 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
      841 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
      901 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
      961 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     1021 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1741 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1801 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aggggtgaca
     1861 catgtttgcc aattgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
     1921 tccagcctta aggataaata acttcgtata ggatacttta tacgaagtta tggcaaagat
     1981 aactcgaagc cgcggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc
     2041 acctcaccca tcaattttat aaaatttgag ctccaccgcg gtggcggccg ctctagaact
     2101 agtggatccc ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag
     2161 ggggggcccg gtaccaattt attttaatta tctacccact ccacctcatg cgcgggcccc
     2221 tcgggttccc gtgcgggacc gtgggcgtgg cgccgaagct cacagacact gctggtataa
     2281 cttcgtatag catacattat acgaagttat gcgatatccc ctatagtgag tcgtatta
//
