LOCUS       sequence-37605-e        2746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2746)
  AUTHORS   Sanjana NE, Cong L, Zhou Y, Cunniff MM, Feng G, Zhang F
  TITLE     A transcription activator-like effector toolbox for genome 
            engineering.
  JOURNAL   Nat Protoc. 2012 Jan 5;7(1):171-92. doi: 10.1038/nprot.2011.431.
  PUBMED    22222791
REFERENCE   2  (bases 1 to 2746)
  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"; doi: 
            "10.1038/nprot.2011"; journalName: "Nat Protoc"; date: "2012-01-5- 
            5"; volume: "7"; issue: "1"; pages: "171-92"
COMMENT     Sequence Label: pNI_v2
FEATURES             Location/Qualifiers
     source          1..2746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(810..901)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     968..987
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1748..1765)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1748..1764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1863..1880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2034..2622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2114..2133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2746,1..809))
                     /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"
ORIGIN
        1 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
       61 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
      121 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
      181 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
      241 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
      301 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
      361 gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgag
      421 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
      481 tcttctaata cctggaacgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
      541 tcaggagtac ggataaaatg cttgatggtc ggaagtggca taaattccgt cagccagttt
      601 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
      661 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
      721 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
      781 ctcgacgttt cccgttgaat atggctcata ttcttccttt ttcaatatta ttgaagcatt
      841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
      901 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
      961 cctgaatatg gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc
     1021 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc
     1081 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1141 gggcctttcg cccgggctaa ttagggggtg tcgcccttcg ctgaactcac cccagagcag
     1201 gtcgtggcaa ttgcgagcaa catcggggga aagcaggcac tcgaaaccgt ccagaggttg
     1261 ctgcctgtgc tgtgccaagc gcacggacgt caaaagggcg acacaaaatt tattctaaat
     1321 gcataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     1381 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     1441 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     1501 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     1561 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     1621 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     1681 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     1741 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     1801 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     1861 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1921 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1981 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2041 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2101 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2161 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2221 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2281 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2341 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2401 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     2461 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2521 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2581 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2641 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     2701 tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc tgcttt
//
