LOCUS       pNHT7::RPL15::sGluc        2993 bp    DNA     circular UNA 10-JUN-2022
DEFINITION  Ligation of 2 sequences.
ACCESSION   urn.local...m-euweyk6
VERSION     urn.local...m-euweyk6
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          join(1..>553,<1258..2993)
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     terminator      268..295
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /label="rrnB T2 terminator"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /label="Escherichia coli rrnB terminator"
     primer_bind     537..553
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 fwd primer bind"
     misc_feature    <550..553
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        550..553
                     /label="Ligation"
     source          join(complement(<1252..>1261),complement(<550..>572))
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
                     /label="source Cloning vector pUC19"
     source          <550..>553
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     promoter        554..572
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     source          complement(<554..>572)
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     misc_feature    569..572
                     /created_by="richardhall"
                     /label="T1"
     ligation        569..572
                     /label="Ligation"
     5'UTR           573..659
                     /created_by="nhall"
                     /label="RPL15 5' UTR"
     CDS             660..>1218
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /note="human codon-optimized"
                     /Original_Translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNF
                     ATTDLDADRGKLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGR
                     CHTYEGDKESAQGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANV
                     QCSDLLKKWLPQRCATFASKIQGQVDKIKGAGGD"
                     /modified_by="nhall"
                     /label="sGluc"
     misc_feature    660..1218
                     /Source="referenceToDocument: urn:local:.:4s-dtuie1k"
                     /label="sGluc (digested by BsaI) Fragment 2 (concatenated
                     sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    660..>663
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        660..663
                     /label="Ligation"
     source          <660..>1218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     misc_feature    1215..663
                     /Source="referenceToDocument: urn:local:.:25-dtuhdjr"
                     /label="Digestion of pNHT7::RPL15 extraction - Fragment 2
                     (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    <1215..1218
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        1215..1218
                     /label="Ligation"
     misc_feature    1218
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     source          1218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     3'UTR           1219..1251
                     /created_by="nhall"
                     /label="RPL15 3' UTR"
     misc_feature    1252..1257
                     /created_by="richardhall"
                     /label="HindIII"
     misc_feature    <1252..>1261
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        1252..1255
                     /label="Ligation"
     source          complement(<1252..>1257)
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
                     /label="source Cloning vector pUC19"
     primer_bind     complement(1258..1274)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 rev primer bind"
     ligation        1258..1261
                     /label="Ligation"
     source          <1258..>1261
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     CDS             1387..2196
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418
                      (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
                     /label="aph(3')-Ia CDS"
     rep_origin      2343..2931
                     /direction="RIGHT"
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori rep origin"
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 tgggtgtgaa agtattattt gcattgatct gtatcgccgt tgcagaagct aaacctactg
      721 aaaataacga ggactttaac attgttgctg tcgctagtaa ttttgctacc actgatctgg
      781 atgccgatag aggaaaactt cctggaaaga aattgccgtt agaagtattg aaagaaatgg
      841 aagctaatgc cagaaaggct ggatgcaccc gaggttgctt aatctgttta agtcatataa
      901 aatgtacacc gaaaatgaag aaatttatac ctggtagatg tcatacttat gagggagata
      961 aagaaagtgc tcagggtgga attggagaag ctattgttga tattccagaa attcctggat
     1021 ttaaagattt ggaacctatg gaacaattta tcgcccaagt tgatctctgt gttgactgta
     1081 caaccggatg cctgaagggc ctggcaaacg tacaatgctc cgatttactg aaaaagtggt
     1141 taccacagcg atgcgccaca tttgctagta aaatccaagg acaggttgat aagataaagg
     1201 gagcaggagg ggattaatga 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 tcgtctcgct 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
//
