LOCUS       sequence-491505-        2326 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Multi-Janus spacer for Galloway Lab Golden Gate cloning scheme. Used
            as a ""filler"" plasmid in transfection experiments..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2326)
  AUTHORS   Kabaria SR, Bae Y, Ehmann ME, Lende-Dorn BA, Beitz AM, Peterman EL, 
            Love KS, Ploessl DS, Galloway KE
  TITLE     Programmable promoter editing for precise control of transgene 
            expression.
  JOURNAL   Nat Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y.
  PUBMED    41083901
REFERENCE   2  (bases 1 to 2326)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y."
COMMENT     Sequence Label: Multi-Janus-Spacer-1x_pKG1126
FEATURES             Location/Qualifiers
     source          1..2326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(116..135)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             127..936
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(194..213)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1026..1614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1515..1534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1779..1865
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1957..1984
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1992..2015
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     primer_bind     complement(2214..2233)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 tgatatctct tagactgcag gtggcccgtg tctcaaaatc tctgatgtta cattgcacaa
       61 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
      121 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
      181 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
      241 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
      301 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
      361 cttccgacca tcaagcattt tatccgtact cctggtgatg catggttact caccactgcg
      421 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
      481 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
      541 tttaacagcg atcgcgtatt tcgtctagct caggcgcaat cacgaatgaa taacggtttg
      601 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
      661 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
      721 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
      781 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
      841 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
      901 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
      961 cattattcag attgggcccc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1021 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1081 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1141 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1201 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1261 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1321 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1381 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1441 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1501 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1561 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1621 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1681 cctgcgttat cccctgattc tgtggataac cgtattaccg ctagcatgga tctcggggac
     1741 gtctaactac taagcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1801 cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1861 caaatccgcc gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag
     1921 gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc
     1981 tttttgcgtt tctacaaact cttcctgtta gttagttact taagctcggg ccccacctgc
     2041 agtccgaaca tggtagctag tggatccttc gtcgacctgc agactggctg tgtataaggg
     2101 agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc
     2161 ggtggctgag atcagccact tcttccccga taacggagac cggcacactg gcgcatatcg
     2221 gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag ttcacgcata
     2281 tgaagctgga catcacctcc cacaacctcg aggtggaaca gtacgc
//
