LOCUS       V016287                 8192 bp    DNA     circular SYN 20-JUL-2021
DEFINITION  Exported.
ACCESSION   V016287
VERSION     V016287
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8192)
  AUTHORS   Chen V, Griffin ME, Maguin P, Varble A, Hang HC.
  TITLE     RecT recombinase expression enables efficient gene editing in
            Enterococcus
  JOURNAL   Appl Environ Microbiol, AEM0084421 (2021) In press
   PUBMED   34232061
REFERENCE   2  (bases 1 to 8192)
  AUTHORS   Chen V, Griffin ME, Hang HC.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAR-2021) Laboratory of Chemical Biology and Microbial
            Pathogenesis, The Rockefeller University, 1230 York Ave., New York,
            NY 10065, USA
REFERENCE   3  (bases 1 to 8192)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Appl
            Environ Microbiol, AEM0084421 (2021) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-MAR-2021) Laboratory of Chemical Biology and Microbial
            Pathogenesis, The Rockefeller University, 1230 York Ave., New York,
            NY 10065, USA"
FEATURES             Location/Qualifiers
     source          1..8192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /label="AmpR promoter"
     CDS             599..1456
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1630..2218
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2404..2546)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2651..2839)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3614..3635)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3651..4730)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4731..4808)
                     /label="lacI promoter"
     promoter        5117..5135
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5136..5160
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5175..5197
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    5204..6247
                     /label="homology arm 1"
                     /note="homology arm 1"
     gene            5204..6193
                     /gene="CVT45_04745"
                     /label="CVT45_04745"
     gene            6204..6247
                     /gene="thyA"
                     /label="thyA"
     regulatory      6278..6330
                     /regulatory_class="promoter"
     regulatory      6288..6293
                     /regulatory_class="minus_35_signal"
     regulatory      6312..6317
                     /regulatory_class="minus_10_signal"
     regulatory      6348..6353
                     /regulatory_class="ribosome_binding_site"
     CDS             6360..7007
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    7035..8034
                     /label="homology arm 2"
                     /note="homology arm 2"
     gene            7035..7938
                     /gene="thyA"
                     /label="thyA"
     gene            7958..8035
                     /gene="CVT45_04755"
                     /label="CVT45_04755"
     CDS             8036..8053
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      8120..8167
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     3901 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     5161 cctctagaaa taattttgtt taactttaag aaggagatat acatatatca agtaaaccaa
     5221 gaagatgact tagaattgaa tctagcgaca caacgtttag gaaaaaaagt actggaaatc
     5281 aaagatggca gttaccgtat aaatgaccaa acactgctag aacatttaga tttgctgatt
     5341 caatctcggg aaaggctagg aattactggt aaaaatgggg cagggaaatc gacattgctt
     5401 aatattctag ctggcagaat tcctctgggt agcggtacaa tttctatcgg tgaaacggtt
     5461 cggcttgcgt attatacaca agaaaatgaa gaaatggcac cagataaacg gatgattgct
     5521 tacttgcaag aagctgctga agaagcaaaa acggctgatg gcagtcaaat cggggtagca
     5581 gaattacttg aacgattttt atttccgaga tttatgcatg ggacactgat tggcaaactt
     5641 tcaggcggag aaaaacggcg attgtttttg ttgaaactgc tgatccaaca acccaatgtg
     5701 ttattattgg atgaaccgac caatgatttg gacatagcaa cattgacgat tttggaagac
     5761 tatttcagaa gtttcccagg tgctgtaatc actgtttccc atgaccgtta tttcttggat
     5821 aaggtagcag ataagctatt ggtattccaa ggaaacggaa aacaagaact ttattatgga
     5881 aacatgtcca gttatttgct gaagcaaaaa gaactccagc agcctgctga aaaagtgaag
     5941 ccaaaaacac agagcaagga acctgctgga aagaaaaaat tatcttatat ggagcaaaaa
     6001 gaatgggaaa cgattgaaga ggatattgca gaacttgaag aaaaaatctc attgctccaa
     6061 gaagagatga atcatcaagg tgatgatttc actcggttac aggaattgca aaacgacgta
     6121 tctgagacag aagcacaact tgaagaaaaa atggcacgtt gggaatattt gagtgaatgg
     6181 gcagaagatt aggaggaaaa gggatggaac aagcgtatct tgacttagga agaaaattgt
     6241 tggaagaaaa aaaaatagat tgaaactcaa gatagatatg ttattgcttg catcaaaata
     6301 aactacatgg gtataatagc aatgaaatgc atttcaaaaa tattttgagg aggagtatca
     6361 tgaactttaa taaaattgat ttagacaatt ggaagagaaa agagatattt aatcattatt
     6421 tgaaccaaca aacgactttt agtataacca cagaaattga tattagtgtt ttataccgaa
     6481 acataaaaca agaaggatat aaattttacc ctgcatttat tttcttagtg acaagggtga
     6541 taaactcaaa tacagctttt agaactggtt acaatagcga cggagagtta ggttattggg
     6601 ataagttaga gccactttat acaatttttg atggtgtatc taaaacattc tctggtattt
     6661 ggactcctgt aaagaatgac ttcaaagagt tttatgattt atacctttct gatgtagaga
     6721 aatataatgg ttcggggaaa ttgtttccca aaacacctat acctgaaaat gctttttctc
     6781 tttctattat tccatggact tcatttactg ggtttaactt aaatatcaat aataatagta
     6841 attaccttct acccattatt acagcaggaa aattcattaa taaaggtaat tcaatatatt
     6901 taccgctatc tttacaggta catcattctg tttgtgatgg ttatcatgca ggattgttta
     6961 tgaactctat tcaggaattg tcagataggc ctaatgactg gcttttataa ttttttgagt
     7021 gatcgttaaa tttaaggaca tcgtaaagaa gacagaacgg gcacgggtac gttaagttta
     7081 ttcgggtacc aaatgcggtt tgatctgcaa aaagggtttc ctttgttaac aacgaaacga
     7141 gtaccttttg gcttgatcaa aagcgaactg ctttggtttt taaaaggaga tacgaacatc
     7201 cgctatctat tacaaaataa aaaccatatt tgggatgagt gggcatttga acgctatatc
     7261 aaaagtacag attataacgg accagatatg actgattttg gacatagaag tttggcagat
     7321 cctgaattca atgaagaata caaaaagcag agcaaacttt tttgcgaaaa gatcctaaca
     7381 gatgattcct ttgctgagaa atacggtgat cttggtcaca tctacggtta tcaatggcgg
     7441 cattgggaga caaaagacgg cggttttatc gatcaaatca aagaagtgat cgaagcaatc
     7501 aaaaagacac ctgattctag aagattgatt gtctctgcat ggaatccaga agatgttcct
     7561 agcatggctt tgcctccttg ccatacaatg tttcaatttt atgtgcaaga gggacggtta
     7621 agctgccaat tgtatcaacg tagtggggat gtctttctag gtgttccgtt caatattgca
     7681 agttatgcgt tgctgactca cttgattgca catgaaacag gtttagaggt gggagaattt
     7741 gttcatacat taggagatgc tcatctttat atcaaccata tcgaacaaat gaaagaacaa
     7801 ttaagccgag aagtacgcgc atttccaact ttagctttga atcaggaaaa gcagtcagta
     7861 tttgattttg agatggaaga tatccagtta gaaggatatg acccacatcc aacaatcaaa
     7921 gcgcctatcg ctgtttaaaa tcagaaagga tgagaaaatg ttagctgcaa tttgggcaca
     7981 ggacgaaaac ggattgattg gaaaagaaga ccgccttcct tggcgtttgc caaagcacca
     8041 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     8101 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     8161 ttttttgctg aaaggaggaa ctatatccgg at
//
