LOCUS       sequence-375142-        2951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An mRNA production vector encoding human codon-optimized 
            nanoluciferase (hNluc) with RPL15 5' and 3' UTRs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2951)
  AUTHORS   Hall RN, Weill U, Drees L, Leal-Ortiz S, Li H, Khariton M, Chai C, 
            Xue Y, Rosental B, Quake SR, Sanchez Alvarado A, Melosh NA, Fire AZ,
            Rink JC, Wang B
  TITLE     Heterologous reporter expression in the planarian Schmidtea 
            mediterranea through somatic mRNA transfection.
  JOURNAL   Cell Rep Methods. 2022 Sep 20;2(10):100298. doi: 
            10.1016/j.crmeth.2022.100298. eCollection 2022 Oct 24.
  PUBMED    36313809
REFERENCE   2  (bases 1 to 2951)
  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: "Cell Rep 
            Methods."; date: "2022-09-20"; volume: "2(10):100298. doi"; pages: "
            10.1016/j.crmeth.2022.100298. eCollection 2022 Oct 24"
COMMENT     Sequence Label: pNHT7::RPL15::hNluc
FEATURES             Location/Qualifiers
     source          1..2951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        554..572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             660..1175
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(1216..1232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1216..1232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1334..1353)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1345..2154
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1412..1431)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2301..2889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2790..2809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agttaatacg actcactata ggattttact aaaatcaaaa ttaattatcg
      601 gtaaacttaa actttgttta aataaataat ttcaaaatta taaaaaatat ttaaaaacaa
      661 tggtcttcac actcgaagat ttcgttgggg actggcgaca gacagccggc tacaacctgg
      721 accaagtcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa
      781 ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac atccatgtca
      841 tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa atttttaagg
      901 tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc acactggtaa
      961 tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa ggcatcgccg
     1021 tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg
     1081 acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc aacggagtga
     1141 ccggctggcg gctgtgcgaa cgcattctgg cgtaatgact aatattaaat aaaactaatt
     1201 ggaattgtta agcttgtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     1261 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     1321 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     1381 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1441 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     1501 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1561 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1621 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     1681 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1741 cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca ggcgcaatca
     1801 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1861 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     1921 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1981 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     2041 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     2101 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     2161 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     2221 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     2281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2941 gctcacatgt t
//
