LOCUS       sequence-375146-        3186 bp    DNA     circular SYN 15-APR-2025
DEFINITION  An mRNA production vector encoding planarian codon-optimized sNluc2 
            with UMSBP 5' and 3' UTRs beginning at a start codon further 
            downstream of the endogenous gene's start codon...
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3186)
  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 3186)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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::UMSBP::sNluc2_ORF2
FEATURES             Location/Qualifiers
     source          1..3186
                     /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(1451..1467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1451..1467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1569..1588)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1580..2389
                     /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(1647..1666)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2536..3124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3025..3044
                     /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 cccgactggg 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 gggtttttat gcaaaattta attatttaaa
      601 aatattgata aaattaatat aatattttat tcttataatt acattaaaaa ataaaatcct
      661 gtttgtttgt atacgcattg gaattcacat atttgattaa tcacgaaaaa tgagttcaag
      721 aagaatggag tgttacaatt gtggtaaaga tggtgtttac tttggaagat tttgttggag
      781 attggagaca aactgctggt tacaatctgg atcaggtact ggaacaaggc ggtgttagtt
      841 cattattcca aaacctgggt gtgagtgtaa ctccgattca gcgaatagtg ttgtctggag
      901 aaaatgggct gaagattgat atacacgtca taattccata cgaaggctta agcggtgatc
      961 aaatgggaca aattgaaaaa atttttaaag tagtttaccc agttgacgac catcatttta
     1021 aagttatcct tcattacggt acactggtta tagatggtgt aactccaaat atgatcgatt
     1081 atttcggaag accttacgaa ggcatagccg tttttgatgg aaaaaagatt acagtaacag
     1141 gtacattgtg gaacggaaat aagattattg acgaacgttt aattaaccca gatggaagtt
     1201 tgctctttag agttacaatt aatggtgtga caggatggag attatgcgaa cggatactcg
     1261 cgtaatgaga taaagcattt tttgtatata tgagcaataa atgtttttaa aataagtttt
     1321 agcgttttcg gtgttaattg gttgtattga ctgtactttt attaataagt agatttatgt
     1381 actatttata ataaattctt gtatcagtct ttagttattt cattgtaaat acaatttaaa
     1441 gtgtaagctt gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1501 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1561 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1621 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1681 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1741 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1801 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1861 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1921 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1981 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat
     2041 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2101 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2161 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2221 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2281 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2341 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2401 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2461 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2521 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2581 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2641 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2701 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2761 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2821 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2881 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2941 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3001 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3061 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3121 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3181 catgtt
//
