LOCUS       sequence-465509-        3128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for making modRNA encoding mGreenLantern.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3128)
  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 3128)
  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-mGL
FEATURES             Location/Qualifiers
     source          1..3128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      6..15
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(12..14,15..17,18..731)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(57..78)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    833..1227
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(1512..1531)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1523..2332
                     /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(1590..1609)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2422..3010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2911..2930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatgagccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
       61 tcgagctgga cggcgacgta aacggccaca agttcagcgt ccgcggcgag ggcgagggcg
      121 atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      181 cctggcccac cctcgtgacc accttaggct acggcgtggc ctgcttcgcc cgctaccccg
      241 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      301 gcaccatctc tttcaaggac gacggtacct acaagacccg cgccgaggtg aagttcgagg
      361 gcgacaccct ggtgaaccgc atcgtgctga agggcatcga cttcaaggag gacggcaaca
      421 tcctggggca caagctggag tacaacttca acagccacaa ggtctatatc acggccgaca
      481 agcagaagaa cggcatcaag gctaacttca agacccgcca caacgttgag gacggcggcg
      541 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      601 ccgacaacca ctacctgagc catcagtcca aactgagcaa agaccccaac gagaagcgcg
      661 atcacatggt cctgaaggag agggtgaccg ccgccgggat tacacatgac atggacgagc
      721 tgtacaagta acaacctcga gctcaagctt cgaattcacc ccaccagtgc aggctgccta
      781 tcagaaagtg gtggctggtg tggctaatgc cctggcccac aagtatcact aagctcgctt
      841 tcttgctgtc caatttctat taaaggttcc tttgttccct aagtccaact actaaactgg
      901 gggatattat gaagggcctt gagcatctgg attctgccta ataaaaaaca tttattttca
      961 ttgcaatgat gtatttaaat tatttctgaa tattttacta aaaagggaat gtgggaggtc
     1021 agtgcattta aaacataaag aaatgaagag ctagttcaaa ccttgggaaa atacactata
     1081 tcttaaactc catgaaagaa ggtgaggctg caaacagcta atgcacattg gcaacagccc
     1141 ctgatgccta tgccttattc atccctcaga aaaggattca agtagaggct tgatttggag
     1201 gttaaagttt tgctatgctg tattttaccg aatcggattt tgtttttaat atttcaaaaa
     1261 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1321 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaacgcg
     1381 tcgaagggaa ttaattcttg aagacgaaag ggccaggtgg cacgtgtctc aaaatctctg
     1441 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1501 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1561 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1621 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1681 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1741 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg gtgatgcatg
     1801 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1861 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1921 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctagctcagg cgcaatcacg
     1981 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2041 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2101 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2161 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2221 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2281 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2341 ggttaattgg ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt
     2401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctaat acgactcact
     3061 ataaggctcc tgggcaacgt gctggtctgt gtgctggccc atcactttgg caaagcacgc
     3121 taccggtc
//
