LOCUS       sequence-375137-        2993 bp    DNA     circular SYN 14-APR-2025
DEFINITION  An mRNA production vector encoding human codon-optimized gaussia 
            luciferase (hGluc) with RPL15 5' and 3' UTRs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2993)
  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 2993)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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::hGluc
FEATURES             Location/Qualifiers
     source          1..2993
                     /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..1217
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     primer_bind     complement(1258..1274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1258..1274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1376..1395)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1387..2196
                     /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(1454..1473)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2343..2931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2832..2851
                     /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 tgggagtcaa agttctgttt gccctgatct gcatcgctgt ggccgaggcc aagcccaccg
      721 agaacaacga agacttcaac atcgtggccg tggccagcaa cttcgcgacc acggatctcg
      781 atgctgaccg cgggaagttg cccggcaaga agctgccgct ggaggtgctc aaagagatgg
      841 aagccaatgc ccggaaagct ggctgcacca ggggctgtct gatctgcctg tcccacatca
      901 agtgcacgcc caagatgaag aagttcatcc caggacgctg ccacacctac gaaggcgaca
      961 aagagtccgc acagggcggc ataggcgagg cgatcgtcga cattcctgag attcctgggt
     1021 tcaaggactt ggagcccatg gagcagttca tcgcacaggt cgatctgtgt gtggactgca
     1081 caactggctg cctcaaaggg cttgccaacg tgcagtgttc tgacctgctc aagaagtggc
     1141 tgccgcaacg ctgtgcgacc tttgccagca agatccaggg ccaggtggac aagatcaagg
     1201 gggccggtgg tgactaatga ctaatattaa ataaaactaa ttggaattgt taagcttgtc
     1261 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     1321 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     1381 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     1441 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1501 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1561 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1621 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1681 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1741 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1801 tttaacagcg atcgcgtatt tcgcctcgct caggcgcaat cacgaatgaa taacggtttg
     1861 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1921 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     1981 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2041 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2101 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2161 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     2221 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     2281 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2461 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2881 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2941 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
