LOCUS       pNHT7::UMSBP::sNluc2_ORF1        3152 bp    DNA     circular UNA 24-MAY-2022
DEFINITION  Concatenation of 2 sequences.
ACCESSION   urn.local...12-euwf2hr
VERSION     urn.local...12-euwf2hr
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          join(1..>553,<1417..3152)
                     /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"
     misc_feature    join(550..>572,<1411..1420)
                     /Source="referenceToDocument: urn:local:.:1s-euwf2hs"
                     /label="Digestion of Blue/White Insert (reversed) -
                     Fragment 2 (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     ligation        550..553
                     /label="Ligation"
     source          join(complement(<1411..>1420),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"
     misc_feature    569..1414
                     /Source="referenceToDocument: urn:local:.:1t-euwf2hs"
                     /label="Digestion of UMSBP Flanks - Fragment 2
                     (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     ligation        569..572
                     /label="Ligation"
     5'UTR           573..709
                     /created_by="nhall"
                     /label="UMSBP 5' UTR"
     CDS             710..1222
                     /created_by="richardhall"
                     /label="UMSBP"
     3'UTR           1223..1410
                     /created_by="nhall"
                     /label="UMSBP 3' UTR"
     primer_bind     complement(1223..1242)
                     /created_by="nhall"
                     /Sequence="GCTTTATCTCCTAGATAACT"
                     /Tm="48.7"
                     /%GC="35.0"
                     /Hairpin_Tm="44.3"
                     /Self_Dimer_Tm="8.5"
                     /modified_by="nhall"
                     /label="UMSBP Primer 5"
     primer_bind     complement(1258..1282)
                     /created_by="nhall"
                     /Sequence="CTTATTTTAAAAACATTTATTGCTC"
                     /Tm="50.6"
                     /%GC="20.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="4.4"
                     /modified_by="nhall"
                     /label="UMSBP Primer 4"
     primer_bind     complement(1295..1314)
                     /created_by="nhall"
                     /Sequence="AATACAACCAATTAACACCG"
                     /Tm="51.7"
                     /%GC="35.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /modified_by="nhall"
                     /label="UMSBP Primer 3"
     primer_bind     complement(1336..1363)
                     /created_by="nhall"
                     /Sequence="AATTTATTATAAATAGTACATAAATCTA"
                     /Tm="48.0"
                     /%GC="10.7"
                     /Hairpin_Tm="30.2"
                     /Self_Dimer_Tm="10.2"
                     /modified_by="nhall"
                     /label="UMSBP Primer 2"
     primer_bind     complement(1387..1410)
                     /Sequence="ACACTTTAAATTGTATTTACAATG"
                     /Tm="50.5"
                     /%GC="20.8"
                     /Hairpin_Tm="51.3"
                     /Self_Dimer_Tm="14.9"
                     /created_by="nhall"
                     /label="UMSBP 3' Primer 1"
     misc_feature    <1411..>1420
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    1411..1416
                     /created_by="richardhall"
                     /label="HindIII"
     misc_feature    1411..572
                     /Source="referenceToDocument: urn:local:.:1r-euwf2hs"
                     /label="Digestion of pNHT7 - Fragment 2 (concatenated
                     sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     ligation        1411..1414
                     /label="Ligation"
     source          complement(<1411..>1416)
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
                     /label="source Cloning vector pUC19"
     ligation        1417..1420
                     /label="Ligation"
     source          <1417..>1420
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     primer_bind     complement(1417..1433)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 rev primer bind"
     CDS             1546..2355
                     /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      2502..3090
                     /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 gggtttttat gcaaaattta attatttaaa
      601 aatattgata aaattaatat aatattttat tcttataatt acattaaaaa ataaaatcct
      661 gtttgtttgt atacgcattg gaattcacat atttgattaa tcacgaaaaa tgagttcaag
      721 aagaatggag tgttacaatt gtggtaaaga tggccatatg tcaagagatt gtactgaaag
      781 taagaagtct ggatctggtc cacgatcagg agcttgcttc aattgcggtg aaagtggtca
      841 tatgtctaga gaatgccctc aaaatggcag tcgtggaggc gaccgtagag gaggctatga
      901 acgcagagga gatgatagac gaggaaacca aagaggtact gaatgttata actgtcatga
      961 aagtggacac atgtcgagag aatgtccaaa acaaagaagc aatggcccaa gaagtaatgg
     1021 ctgttataaa tgtggtgaaa gtggccatat ggctcgtgat tgtaaccaag aaggtagtga
     1081 taatagacaa gaaagacaag ttaaatgtta caattgtaat caaaatggcc atatgtcgag
     1141 agaatgcccc aatggacaat cggaaaagaa atgttacaag tgtaacaaaa ttggacatat
     1201 gtccagagag tgcaacgaat aaagttatct aggagataaa gcattttttg tatatatgag
     1261 caataaatgt ttttaaaata agttttagcg ttttcggtgt taattggttg tattgactgt
     1321 acttttatta ataagtagat ttatgtacta tttataataa attcttgtat cagtctttag
     1381 ttatttcatt gtaaatacaa tttaaagtgt aagcttgtca tagctgtttc ctggcagctc
     1441 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1501 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1561 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1621 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1681 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1741 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1801 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     1861 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1921 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1981 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2041 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2101 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2161 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2221 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2281 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2341 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2401 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     2461 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2521 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2581 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2641 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2701 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2761 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2821 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2881 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2941 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3001 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3061 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3121 ctggcctttt gctggccttt tgctcacatg tt
//
