LOCUS       pENTR2b-lifeAct-        3072 bp DNA     circular SYN 11-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3072)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2024 from SnapGene Viewer 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3072
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    458..493
                     /label=MCS 1
                     /note="multiple cloning site, part 1"
     misc_feature    494
                     /label=overhang+Kpn1
     misc_feature    495
                     /label=T changed to G for Kozacs seq restauration
     misc_feature    501..551
                     /label=LifeAct
     misc_feature    573..1283
                     /label=mNeonGreen
     misc_feature    1284..1286
                     /label=2nd stop codon
                     /note="Inserted by me. "
     misc_feature    1287
                     /label=XhoI+overhang
     misc_feature    1288..1297
                     /label=MCS 2
                     /note="multiple cloning site, part 2"
     protein_bind    complement(1301..1400)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1523..2332
                     /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"
     rep_origin      2422..3010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 ctggatccgg taccgccacc atgggcgtgg ccgacctgat caagaagttc gagagcatca
      541 gcaaggagga gggggatcca ccggtcgcca ccatggtgag caagggcgag gaggataaca
      601 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
      661 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
      721 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
      781 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag
      841 atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
      901 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
      961 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
     1021 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
     1081 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
     1141 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
     1201 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
     1261 gcatggacga gctgtacaag taatagctcg agatatctag acccagcttt cttgtacaaa
     1321 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1381 caaaataaaa tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg
     1441 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1501 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1561 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1621 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1681 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1741 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1801 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1861 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1921 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1981 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2041 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2101 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2161 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2221 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2281 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2341 ggttaattgg ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt
     2401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3061 tgctcacatg tt
//
