LOCUS       sequence-465502-        3011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for making modRNA encoding the Puromycin resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3011)
  AUTHORS   Blanch-Asensio A, Ploessl DS, Wang NB, Mummery CL, Galloway KE, 
            Davis RP
  TITLE     STRAIGHT-IN Dual: a platform for dual, single-copy integrations of 
            DNA payloads and gene circuits into human induced pluripotent stem 
            cells.
  JOURNAL   bioRxiv [Preprint]. 2024 Oct 17:2024.10.17.616637. doi: 
            10.1101/2024.10.17.616637.
  PUBMED    39464154
REFERENCE   2  (bases 1 to 3011)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Oct 17:2024.10.17.616637. doi: 
            10.1101/2024.10.17.616637."
COMMENT     Sequence Label: pIVT-PuroR
FEATURES             Location/Qualifiers
     source          1..3011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      180..768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     669..688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             899..1498
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(899..918)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1395..1415
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1600..1994
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2292..3011,1..90)
                     /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(2359..2378)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
       61 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacattat
      121 tcagattggg ccccgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      181 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      241 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      301 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
      361 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      421 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      481 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      541 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      601 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      661 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      721 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      781 gcggcctttt tacggttcct ggcctaatac gactcactat aaggctcctg ggcaacgtgc
      841 tggtctgtgt gctggcccat cactttggca aagcacgcta ccggtcaatg aagccaccat
      901 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
      961 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     1021 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     1081 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     1141 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     1201 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     1261 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     1321 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     1381 gacatccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     1441 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaca
     1501 acctcgagct caagcttcga attcacccca ccagtgcagg ctgcctatca gaaagtggtg
     1561 gctggtgtgg ctaatgccct ggcccacaag tatcactaag ctcgctttct tgctgtccaa
     1621 tttctattaa aggttccttt gttccctaag tccaactact aaactggggg atattatgaa
     1681 gggccttgag catctggatt ctgcctaata aaaaacattt attttcattg caatgatgta
     1741 tttaaattat ttctgaatat tttactaaaa agggaatgtg ggaggtcagt gcatttaaaa
     1801 cataaagaaa tgaagagcta gttcaaacct tgggaaaata cactatatct taaactccat
     1861 gaaagaaggt gaggctgcaa acagctaatg cacattggca acagcccctg atgcctatgc
     1921 cttattcatc cctcagaaaa ggattcaagt agaggcttga tttggaggtt aaagttttgc
     1981 tatgctgtat tttaccgaat cggattttgt ttttaatatt tcaaaaaaaa aaaaaaaaaa
     2041 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2101 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaacgcgt cgaagggaat
     2161 taattcttga agacgaaagg gccaggtggc acgtgtctca aaatctctga tgttacattg
     2221 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2281 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2341 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2401 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2461 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2521 tgcctcttcc gaccatcaag cattttatcc gtactcctgg tgatgcatgg ttactcacca
     2581 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2641 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2701 gtccttttaa cagcgatcgc gtatttcgtc tagctcaggc gcaatcacga atgaataacg
     2761 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2821 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2881 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2941 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3001 tttctccttc a
//
