LOCUS       Exported                5930 bp ds-DNA     circular SYN 18-DEC-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  AUTHORS   Doudna Lab
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 18, 2019 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5930
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3212..3556
                     /organism="Staphylococcus delphini"
                     /host="Equus ferus caballus"
                     /strain="214092504301-1"
                     /mol_type="genomic DNA"
                     /country="Netherlands"
                     /lat_lon="52.5 N 5.75 E"
                     /isolation_source="nasal swab"
                     /collected_by="VMDC"
                     /collection_date=25-Sep-2014
                     /note="color: #ffffff"
                     /db_xref="taxon:53344"
     misc_feature    233..375
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(477..668)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        2229..2247
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     RBS             2336..2358
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             2366..2407
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /note="color: #cc99b2"
                     /translation="MKSSHHHHHHHHHH"
     CDS             2417..3130
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="color: #05fd14"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3185..3205
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /note="color: #cc99b2
                     Cleavage site after base 3202"
                     /translation="ENLYFQS"
     CDS             3212..3556
                     /codon_start=1
                     /locus_tag="B5C05_01750"
                     /product="hypothetical protein"
                     /label=truncated AcrIIA15
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Protein Homology."
                     /note="color: #00ccff"
                     /inference="COORDINATES:same species:RefSeq"
                     /protein_id="PCF60932.1"
                     /translation="MEKVDTWIVYRGRTADMNKSYIAEGSTYEEVYNNFVDKYGYDVLD
                     EDIYEIQLLKKNGENLDDYDVDSDGINNYDKLDEFRESDYVDLEDYDYRELFENSSSQV
                     YYHEFEITHE"
     gene            3212..3556
                     /locus_tag="B5C05_01750"
                     /label=B5C05_01750
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3557..3561
                     /label=Islander Island
                     /note="color: #a6acb3"
     misc_feature    3557..3561
                     /label=Comparator Island
                     /note="color: #00ff00"
     gene            3557..3561
                     /locus_tag="AAX06_RS01960"
                     /old_locus_tag="AAX06_01960"
                     /label=AAX06_RS01960
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3557..3561
                     /label=PHASTER prophage region
                     /note="color: #a6acb3"
     terminator      3699..3746
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     promoter        complement(4112..4140)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
                     /note="This reverse directional feature has 3 segments:
                        1: 4112 .. 4117 / #ffffff / -10
                        2: 4118 .. 4134 / #ffffff
                        3: 4135 .. 4140 / #ffffff / -35"
     promoter        4253..4357
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4358..5218
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4358 .. 4426 / #ccffcc / signal sequence
                        2: 4427 .. 5218 / #ccffcc
                     Cleavage site after base 4426"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      join(5389..5930,1..47)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
       61 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      121 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      181 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
      241 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag
      301 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
      361 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      421 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
      481 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
      541 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
      601 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
      661 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
      721 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
      781 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
      841 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
      901 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
      961 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     1021 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     1081 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     1141 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     1201 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     1261 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     1321 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     1381 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     1441 gcggtccagt gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     1501 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     1561 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     1621 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     1681 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     1741 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     1801 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     1861 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     1921 gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     1981 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     2041 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     2101 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     2161 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     2221 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gtgccggctc
     2281 cggagagctc tttaattaag cggccgccct gcaggactcg agttctagaa ataattttgt
     2341 ttaactttaa gaaggagata tacatatgaa atcttctcac catcaccatc accatcacca
     2401 tcaccatggt tcttctatga gcaaaggaga agaacttttc actggagttg tcccaattct
     2461 tgttgaatta gatggtgatg ttaatgggca caaattttct gtccgtggag agggtgaagg
     2521 tgatgctaca aacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt
     2581 tccgtggcca acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc
     2641 ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     2701 acgcactata tctttcaaag atgacgggac ctacaagacg cgtgctgaag tcaagtttga
     2761 aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag aagatggaaa
     2821 cattcttgga cacaaactcg agtacaactt taactcacac aatgtataca tcacggcaga
     2881 caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacgttg aagatggttc
     2941 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     3001 accagacaac cattacctgt cgacacaatc tgtcctttcg aaagatccca acgaaaagcg
     3061 tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga
     3121 gctctacaaa aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat
     3181 cgaggaaaac ctgtacttcc aatccaatgc aatggaaaaa gtggacacat ggattgttta
     3241 cagaggaaga acagcggata tgaataaaag ctatatagcg gaaggcagca cttacgaaga
     3301 agtttataat aacttcgttg ataaatacgg ctatgatgtt ttagatgagg atatttacga
     3361 aattcaactt ctaaaaaaga acggagaaaa cttggacgat tatgacgttg attctgacgg
     3421 catcaacaat tatgataaat tggacgagtt cagagaatcg gattacgtgg atctagagga
     3481 ttacgattat agagaattgt tcgaaaatag cagctctcaa gtttattatc atgaatttga
     3541 gataacacac gaataataat aacattggaa gtggataacg gatccgcgat cgcggcgcgc
     3601 cacctggtgg ccggccggta ccacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa
     3661 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3721 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg
     3781 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg
     3841 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc
     3901 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg
     3961 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
     4021 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
     4081 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
     4141 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4201 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4321 ctgataaatg cttcaataac attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4381 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4441 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4501 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4561 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     4621 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4681 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4741 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4801 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4861 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     4921 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4981 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5041 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5101 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5161 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5221 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5281 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5521 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
//
