LOCUS       sequence-465504-        3972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for making modRNA encoding mutant Bxb1 modRNA (I87L).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3972)
  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 3972)
  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
FEATURES             Location/Qualifiers
     source          1..3972
                     /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             906..926
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             927..953
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2561..2955
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(3242..3261)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(3253..3972,1..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(3320..3339)
                     /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 cgacccgcca
      901 ccatgccaaa aaagaaaaga aaagtgtatc cctatgatgt ccccgattat gccggttcaa
      961 gagccctggt cgtgattaga ctgagccgag tgacagacgc caccacaagt cccgagagac
     1021 agctggaatc atgccagcag ctctgtgctc agcggggttg ggatgtggtc ggcgtggcag
     1081 aggatctgga cgtgagcggg gccgtcgatc cattcgacag aaagaggagg cccaacctgg
     1141 caagatggct cgctttcgag gaacagccct ttgatgtgat cgtcgcctac agagtggacc
     1201 ggctgacccg ctcactgcga catctccagc agctggtgca ttgggctgag gaccacaaga
     1261 aactggtggt cagcgcaaca gaagcccact tcgatactac cacacctttt gccgctgtgg
     1321 tcatcgcact gatgggcact gtggcccaga tggagctcga agctatcaag gagcgaaaca
     1381 ggagcgcagc ccatttcaat attagggccg gtaaatacag aggctccctg cccccttggg
     1441 gatatctccc taccagggtg gatggggagt ggagactggt gccagacccc gtccagagag
     1501 agcggattct ggaagtgtac cacagagtgg tcgataacca cgaaccactc catctggtgg
     1561 cacacgacct gaatagacgc ggcgtgctct ctccaaagga ttattttgct cagctgcagg
     1621 gaagagagcc acagggaaga gaatggagtg ctactgcact gaagagatct atgatcagtg
     1681 aggctatgct gggttacgca acactcaatg gcaaaactgt ccgggacgat gacggagccc
     1741 ctctggtgag ggctgagcct attctcacca gagagcagct cgaagctctg cgggcagaac
     1801 tggtcaagac tagtcgcgcc aaacctgccg tgagcacccc aagcctgctc ctgagggtgc
     1861 tgttctgcgc cgtctgtgga gagccagcat acaagtttgc cggcggaggg cgcaaacatc
     1921 cccgctatcg atgcaggagc atggggttcc ctaagcactg tggaaacggg acagtggcca
     1981 tggctgagtg ggacgccttt tgcgaggaac aggtgctgga tctcctgggt gacgctgagc
     2041 ggctggaaaa agtgtgggtg gcaggatctg actccgctgt ggagctggca gaagtcaatg
     2101 ccgagctcgt ggatctgact tccctcatcg gatctcctgc atatagagct gggtccccac
     2161 agagagaagc tctggacgca cgaattgctg cactcgctgc tagacaggag gaactggagg
     2221 gcctggaggc caggccctct ggatgggagt ggcgagaaac cggacagagg tttggggatt
     2281 ggtggaggga gcaggacacc gcagccaaga acacatggct gagatccatg aatgtccggc
     2341 tcacattcga cgtgcgcggt ggcctgactc gaaccatcga ttttggcgac ctgcaggagt
     2401 atgaacagca cctgagactg gggtccgtgg tcgaaagact gcacactggg atgtcctagc
     2461 aacctcgagc tcaagcttcg aattcacccc accagtgcag gctgcctatc agaaagtggt
     2521 ggctggtgtg gctaatgccc tggcccacaa gtatcactaa gctcgctttc ttgctgtcca
     2581 atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga
     2641 agggccttga gcatctggat tctgcctaat aaaaaacatt tattttcatt gcaatgatgt
     2701 atttaaatta tttctgaata ttttactaaa aagggaatgt gggaggtcag tgcatttaaa
     2761 acataaagaa atgaagagct agttcaaacc ttgggaaaat acactatatc ttaaactcca
     2821 tgaaagaagg tgaggctgca aacagctaat gcacattggc aacagcccct gatgcctatg
     2881 ccttattcat ccctcagaaa aggattcaag tagaggcttg atttggaggt taaagttttg
     2941 ctatgctgta ttttaccgaa tcggattttg tttttaatat ttcaaaaaaa aaaaaaaaaa
     3001 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3061 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaacgcg tcgaagggaa
     3121 ttaattcttg aagacgaaag ggccaggtgg cacgtgtctc aaaatctctg atgttacatt
     3181 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     3241 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     3301 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     3361 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     3421 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     3481 atgcctcttc cgaccatcaa gcattttatc cgtactcctg gtgatgcatg gttactcacc
     3541 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     3601 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     3661 tgtcctttta acagcgatcg cgtatttcgt ctagctcagg cgcaatcacg aatgaataac
     3721 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3781 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3841 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3901 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3961 ttttctcctt ca
//
