LOCUS       Exported                3402 bp ds-DNA     circular SYN 23-OCT-2020
DEFINITION  B5r-Venus-ns-B1r.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3402)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 29, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     Insert Molecule:   C512-P14, 774 bps DNA (attB5r, attB1r sites)
            Donor Vector:   pDonr-235, 4749 bps DNA circular (attP5r, attP1r 
            sites)
            Recombination using BP clonase to produce Entry Clone
FEATURES             Location/Qualifiers
     source          1..3402
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(43..166)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             171..887
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus-
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    888..1012
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1085..1103)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1108..1124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1549..2340
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2433..3021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3156..3183
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      3275..3361
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcgaattct acaggtcact aataccatct aagtagttga ttcatagtga ctgcatatgt
       61 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
      121 tttatatcat tttacgtttc tcgttcaact tttgtataca aagttgaacc atggtgagca
      181 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      241 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      301 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      361 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      421 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      481 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      541 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      601 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
      661 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
      721 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
      781 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      841 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaca actttgtaca
      901 aaaaagttga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
      961 ataaaaaaca gactacataa tactgtaaaa cacaacatat gcagtcacta tgaaccaact
     1021 acttagatgg tattagtgac ctgtagaatt cgagctctag agctgcaggg cggccgcgat
     1081 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1141 tctcaaaatc tctgatgtta cattgcacaa gatcggtacc cggggatcct ctagaccagc
     1201 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     1261 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     1321 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     1381 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     1441 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     1501 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     1561 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     1621 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     1681 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     1741 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     1801 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     1861 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     1921 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     1981 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     2041 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     2101 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     2161 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     2221 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     2281 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     2341 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2401 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2461 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2521 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2581 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2641 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2701 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2761 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2821 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2881 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2941 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3001 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3061 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3121 attaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3181 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3241 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3301 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3361 gatgcctggc agttccctac tctcgcgtta acgctaccat gg
//
