LOCUS       Exported                3969 bp ds-DNA     circular SYN 20-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMMDS DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3969)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 21, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3969
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             967..1272
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1313..1437)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1536..1569)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      1571..1927
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             2390..3181
                     /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"
     promoter        3272..3381
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     polyA_signal    3517..3565
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    3789..3913
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccagatc tggatccggc ttactaaaag ccagataaca
      661 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
      721 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
      781 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
      841 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
      901 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
      961 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     1021 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     1081 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     1141 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     1201 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     1261 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     1321 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     1381 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggttga
     1441 taagcttaaa cgtcgaggga tcttcataag agaagaggga cagctatgac tgggagtagt
     1501 caggagagga ggaaaaaaac ccatgtgcct ggcagataac ttcgtataat gtatgctata
     1561 cgaagttatg gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     1621 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     1681 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     1741 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     1801 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     1861 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     1921 gctgacataa ttcagcttca cgctgccgca agcactcagg gcgcaagggc tgctaaagga
     1981 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     2041 gggaggcaga ataaatgatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa
     2101 actggcctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     2161 taagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     2221 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     2281 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2341 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc
     2401 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2461 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     2521 acacagtgat attgatttgc tggttacggt gacggtaagg cttgatgaaa caacgcggcg
     2581 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     2641 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     2701 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     2761 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     2821 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2881 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     2941 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     3001 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     3061 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     3121 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     3181 atgtctaaca attcgttcaa gccgacggat ctatgtcggg tgcggagaaa gaggtaatga
     3241 aatggcacct aggtatcgat actagtatac ggacctttaa ttcaacccaa cacaatatat
     3301 tatagttaaa taagaattat tatcaaatca tttgtatatt aattaaaata ctatactgta
     3361 aattacattt tatttacaat cactcgacga agacttgatc acccgggatc tcgagccatg
     3421 gtgctagcag ctgatgcata gcatgcggta ccgggagatg ggggaggcta actgaaacac
     3481 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3541 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3601 gtcgataccc caccgagacc ccattgggac caatacgccc gcgtttcttc cttttcccca
     3661 ccccaacccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaagc
     3721 cctgccatag ccactacggg tacgtttctg taactataac ggtcctaagg tagcgagttt
     3781 aaacatcaac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
     3841 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     3901 tccagtcact atggcggccg cattaggcca ctgagcgtca gaccccgtag aaaagatcaa
     3961 aggatcttc
//
