LOCUS       sequence-356207-        3160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses kanamycin when ligated in-frame.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3160)
  AUTHORS   Kudo T, Lane K, Covert MW
  TITLE     A multiplexed epitope barcoding strategy that enables dynamic 
            cellular phenotypic screens.
  JOURNAL   Cell Syst. 2022 May 18;13(5):376-387.e8. doi: 
            10.1016/j.cels.2022.02.006. Epub 2022 Mar 21.
  PUBMED    35316656
REFERENCE   2  (bases 1 to 3160)
  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: 
            "10.1016/j.cels.2022.02.006"; journalName: "Cell Syst"; date: 
            "2022-05-18- 18"; volume: "13"; issue: "5"; pages: "376-387.e8"
COMMENT     Sequence Label: in-frame kanamycin backbone
FEATURES             Location/Qualifiers
     source          1..3160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     287..306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      648..1236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1137..1156
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1390..1407
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1783..2574
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1783..2574
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1834..1853)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2444..2463
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(2824..3160,1..455,456..524))
                     /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"
ORIGIN
        1 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
       61 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
      121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
      181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
      241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
      301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
      361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
      421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
      481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
      541 tattattgaa gcatttatca gggttattgt ctcatgacca aaatccctta acgtgagttt
      601 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      661 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      721 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      781 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      841 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      901 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      961 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1021 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1081 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1141 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1201 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1261 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1321 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1381 accgagcgca gcgagtcagt gagcgaggaa gcggaagaca cctagatcct tttcacgtag
     1441 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     1501 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     1561 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     1621 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     1681 tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgggg
     1741 cgcgcctgat catagatcct gcagggcgat cgcgtaatta gcattgaaca agatggattg
     1801 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     1861 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     1921 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     1981 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     2041 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     2101 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     2161 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     2221 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     2281 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     2341 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     2401 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     2461 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     2521 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattatt
     2581 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2641 cgcatcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2701 aatacattca aatatgtatc cgctcatgag attatcaaaa aggatcttca cctagatcct
     2761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2941 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
//
