LOCUS       sequence-465497-        3911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for making modRNA encoding mutant Bxb1modRNA (I87L, 
            A369P, E434G).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3911)
  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 3911)
  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-Bxb1-I87L, A369P, E434G
FEATURES             Location/Qualifiers
     source          1..3911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(11..37)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(38..58)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(874..1683)
                     /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     1597..1616
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1675..1694
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     polyA_signal    1920..2314
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
ORIGIN
        1 ctcttgaacc ggcataatcg gggacatcat agggatacac ttttcttttc ttttttggca
       61 tggtggcggg tcgcattgac cggtagcgtg ctttgccaaa gtgatgggcc agcacacaga
      121 ccagcacgtt gcccaggagc cttatagtga gtcgtattag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg gggcccaatc
      841 tgaataatgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
      901 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
      961 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1021 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1081 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     1141 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1201 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gctagacgaa atacgcgatc
     1261 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     1321 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     1381 tccggggatc gcagtggtga gtaaccatgc atcaccagga gtacggataa aatgcttgat
     1441 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1501 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     1561 caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     1621 taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct
     1681 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     1741 atttttatct tgtgcaatgt aacatcagag attttgagac acgtgccacc tggccctttc
     1801 gtcttcaaga attaattccc ttcgacgcgt tttttttttt tttttttttt tttttttttt
     1861 tttttttttt tttttttttt tttttttttt tgaaatatta aaaacaaaat ccgattcggt
     1921 aaaatacagc atagcaaaac tttaacctcc aaatcaagcc tctacttgaa tccttttctg
     1981 agggatgaat aaggcatagg catcaggggc tgttgccaat gtgcattagc tgtttgcagc
     2041 ctcaccttct ttcatggagt ttaagatata gtgtattttc ccaaggtttg aactagctct
     2101 tcatttcttt atgttttaaa tgcactgacc tcccacattc cctttttagt aaaatattca
     2161 gaaataattt aaatacatca ttgcaatgaa aataaatgtt ttttattagg cagaatccag
     2221 atgctcaagg cccttcataa tatcccccag tttagtagtt ggacttaggg aacaaaggaa
     2281 cctttaatag aaattggaca gcaagaaagc gagcttagtg atacttgtgg gccagggcat
     2341 tagccacacc agccaccact ttctgatagg cagcctgcac tggtggggtg aattcgaagc
     2401 ttgagctcga ggttgctagg acatcccagt gtgcagtctt tcgaccacgg accccagtct
     2461 caggtgctgt tcatactcct gcaggtcgcc aaaatcgatg gttcgagtca ggccaccgcg
     2521 cacgtcgaat gtgagccgga cattcatgga tctcagccat gtgttcttgg ctgcggtgtc
     2581 ctgctccctc caccaatccc caaacctctg tccggtgcct cgccactccc atccagaggg
     2641 cctggcctcc aggccctcca gttcctcctg tctagcagcg agtgcagcaa ttcgtgcgtc
     2701 cagagcttct ctctgtgggg acccagctct atatgcagga gatccgatga gggaagtcag
     2761 atccacgagc tcggcattga cttctgccag ctccacagcg gagtcagatc ctggcaccca
     2821 cactttttcc agccgctcag cgtcacccag gagatccagc acctgttcct cgcaaaaggc
     2881 gtcccactca gccatggcca ctgtcccgtt tccacagtgc ttagggaacc ccatgctcct
     2941 gcatcgatag cggggatgtt tgcgccctcc gccggcaaac ttgtatgctg gctctccaca
     3001 gacggcgcag aacagcaccc tcaggagcag gcttggggtg ctcacggcag gtttggcgcg
     3061 actagtcttg accagttctg cccgcagagc ttcgagctgc tctctggtga gaataggctc
     3121 agccctcacc agaggggctc cgtcatcgtc ccggacagtt ttgccattga gtgttgcgta
     3181 acccagcata gcctcactga tcatagatct cttcagtgca gtagcactcc attctcttcc
     3241 ctgtggctct cttccctgca gctgagcaaa ataatccttt ggagagagca cgccgcgtct
     3301 attcaggtcg tgtgccacca gatggagtgg ttcgtggtta tcgaccactc tgtggtacac
     3361 ttccagaatc cgctctctct ggacggggtc tggcaccagt ctccactccc catccaccct
     3421 ggtagggaga tatccccaag ggggcaggga gcctctgtat ttaccggccc taatattgaa
     3481 atgggctgcg ctcctgtttc gctccttgat agcttcgagc tccatctggg ccacagtgcc
     3541 catcagtgcg atgaccacag cggcaaaagg tgtggtagta tcgaagtggg cttctgttgc
     3601 gctgaccacc agtttcttgt ggtcctcagc ccaatgcacc agctgctgga gatgtcgcag
     3661 tgagcgggtc agccggtcca ctctgtaggc gacgatcaca tcaaagggct gttcctcgaa
     3721 agcgagccat cttgccaggt tgggcctcct ctttctgtcg aatggatcga cggccccgct
     3781 cacgtccaga tcctctgcca cgccgaccac atcccaaccc cgctgagcac agagctgctg
     3841 gcatgattcc agctgtctct cgggacttgt ggtggcgtct gtcactcggc tcagtctaat
     3901 cacgaccagg g
//
