LOCUS       sequence-375098-        2945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An mRNA production vector encoding planarian codon-optimized 
            nanoluciferase (sNluc1) with RPL10 5' and 3' UTRs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2945)
  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 2945)
  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::RPL10::sNluc1
FEATURES             Location/Qualifiers
     source          1..2945
                     /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"
     primer_bind     complement(1210..1226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1210..1226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1328..1347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1339..2148
                     /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(1406..1425)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2295..2883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2784..2803
                     /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 ggctttataa tttttataaa tcattatcaa
      601 ttacccatat atatttgttt aaatacataa atttaatttc catataaaaa tttaaaatgg
      661 tttttacatt agaagatttt gttggagatt ggagacaaac agctggatat aatttagatc
      721 aagttttaga acaaggagga gtttcatcat tgttccaaaa tttgggagtt tcagttacac
      781 caattcaaag aattgttttg agtggagaaa atggattaaa gattgatatt catgttatta
      841 ttccatatga gggattatca ggagatcaaa tgggacaaat tgaaaagatt ttcaaagttg
      901 tttatccagt tgatgatcat catttcaaag ttattttaca ttatggaaca ttagttattg
      961 atggagttac accaaatatg attgattatt ttggaagacc atatgaggga attgctgttt
     1021 ttgatggaaa gaagattaca gttacaggaa cattatggaa tggaaacaaa attattgacg
     1081 aaagattaat taatccagat ggatcattat tatttagagt tacaattaat ggagttacag
     1141 gatggagatt atgtgaaaga attttagctt aatatctttt gaagtgaata aagttttgga
     1201 tttaagcttg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1261 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1321 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1381 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1441 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1501 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1561 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1621 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1681 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1741 tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg
     1801 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1861 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     1921 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1981 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2041 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2101 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     2161 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     2221 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2401 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2521 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2581 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2641 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2701 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2761 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2821 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2881 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2941 atgtt
//
