LOCUS       sequence-375139-        3087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An mRNA production vector encoding planarian codon-optimized 
            nanoluciferase (sNluc2) with enolase 5' and 3' UTRs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3087)
  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 3087)
  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::ENO::sNluc2
FEATURES             Location/Qualifiers
     source          1..3087
                     /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"
     primer_bind     554..573
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        554..572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1352..1368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1352..1368)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1470..1489)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1481..2290
                     /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(1548..1567)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2437..3025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2926..2945
                     /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 gggttttagt aagtattata aatagtaaac
      601 catataaatt gcgtacgtga ttacagaaat tatatttgta atttacaaat tttttcaaga
      661 aaatggtgtt tactttggaa gattttgttg gagattggag acaaactgct ggttacaatc
      721 tggatcaggt actggaacaa ggcggtgtta gttcattatt ccaaaacctg ggtgtgagtg
      781 taactccgat tcagcgaata gtgttgtctg gagaaaatgg gctgaagatt gatatacacg
      841 tcataattcc atacgaaggc ttaagcggtg atcaaatggg acaaattgaa aaaattttta
      901 aagtagttta cccagttgac gaccatcatt ttaaagttat ccttcattac ggtacactgg
      961 ttatagatgg tgtaactcca aatatgatcg attatttcgg aagaccttac gaaggcatag
     1021 ccgtttttga tggaaaaaag attacagtaa caggtacatt gtggaacgga aataagatta
     1081 ttgacgaacg tttaattaac ccagatggaa gtttgctctt tagagttaca attaatggtg
     1141 tgacaggatg gagattatgc gaacggatac tcgcgtaatt tatatatcac tggttttgac
     1201 tctagtattt agtttggcct atattagtgt tgactgacct tgatatactt tttaccggtt
     1261 ttgtgttaat aaatattttt ctccaataat catggattca ttttgatgat tatagctata
     1321 gaaatatata aattttttgg ttgttaagct tgtcatagct gtttcctggc agctctggcc
     1381 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1441 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1501 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1561 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     1621 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1681 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1741 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     1801 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1861 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgcct
     1921 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1981 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2041 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2101 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2161 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2221 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2281 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2341 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2581 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3061 cttttgctgg ccttttgctc acatgtt
//
