LOCUS       sequence-465495-        3369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for making modRNA encoding iCre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3369)
  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 3369)
  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-iCre
FEATURES             Location/Qualifiers
     source          1..3369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(127..195,196..987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGYRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(345..364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1158..1746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1647..1666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1818..1827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1830..1850
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    join(2996..3369,1..21)
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
ORIGIN
        1 gttttgctat gctgtatttt acgcggaacc cctatttgtt tatttttcta aatacattca
       61 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      121 aagagtatga gtattcaaca tttccgtgtc gctcttattc ccttttttgc ggcattttgc
      181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      361 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      841 gggtatcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
      961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1741 tggaaactcc tgggcaacgt gctggtctgt gtgctggccc atcactttgg caaagcacgc
     1801 taccggtcac gcgtgccgcc accatggtgc ccaagaagaa gaggaaagtc tccaacctgc
     1861 tgactgtgca ccaaaacctg cctgccctcc ctgtggatgc cacctctgat gaagtcagga
     1921 agaacctgat ggacatgttc agggacaggc aggccttctc tgaacacacc tggaagatgc
     1981 tcctgtctgt gtgcagatcc tgggctgcct ggtgcaagct gaacaacagg aaatggttcc
     2041 ctgctgaacc tgaggatgtg agggactacc tcctgtacct gcaagccaga ggcctggctg
     2101 tgaagaccat ccaacagcac ctgggccagc tcaacatgct gcacaggaga tctggcctgc
     2161 ctcgcccttc tgactccaat gctgtgtccc tggtgatgag gagaatcaga aaggagaatg
     2221 tggatgctgg ggagagagcc aagcaggccc tggcctttga acgcactgac tttgaccaag
     2281 tcagatccct gatggagaac tctgacagat gccaggacat caggaacctg gccttcctgg
     2341 gcattgccta caacaccctg ctgcgcattg ccgaaattgc cagaatcaga gtgaaggaca
     2401 tctcccgcac cgatggtggg agaatgctga tccacattgg caggaccaag accctggtgt
     2461 ccacagctgg tgtggagaag gccctgtccc tgggggttac caagctggtg gagagatgga
     2521 tctctgtgtc tggtgtggct gatgacccca acaactacct gttctgccgg gtcagaaaga
     2581 atggtgtggc tgccccttct gccacctccc aactgtccac ccgggccctg gaagggatct
     2641 ttgaggccac ccaccgcctg atctatggtg ccaaggatga ctctgggcag agatacctgg
     2701 cctggtctgg ccactctgcc agagtgggtg ctgccaggga catggccagg gctggtgtgt
     2761 ccatccctga aatcatgcag gctggtggct ggaccaatgt gaacattgtg atgaactaca
     2821 tcagaaacct ggactctgag actggggcca tggtgaggct gctcgaggat ggggactgag
     2881 cggccgcact cctcaggtct cgagctcaag cttcgaattc accccaccag tgcaggctgc
     2941 ctatcagaaa gtggtggctg gtgtggctaa tgccctggcc cacaagtatc actaagctcg
     3001 ctttcttgct gtccaatttc tattaaaggt tcctttgttc cctaagtcca actactaaac
     3061 tgggggatat tatgaagggc cttgagcatc tggattctgc ctaataaaaa acatttattt
     3121 tcattgcaat gatgtattta aattatttct gaatatttta ctaaaaaggg aatgtgggag
     3181 gtcagtgcat ttaaaacata aagaaatgaa gagctagttc aaaccttggg aaaatacact
     3241 atatcttaaa ctccatgaaa gaaggtgagg ctgcaaacag ctaatgcaca ttggcaacag
     3301 cccctgatgc ctatgcctta ttcatccctc agaaaaggat tcaagtagag gcttgatttg
     3361 gaggttaaa
//
