LOCUS       KanSacB+Scarlett        5747 bp DNA     circular SYN 08-AUG-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5747)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Dec 20, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5747
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    1..4
                     /label=AATG
                     /note="/color=#f7e8f7"
     misc_feature    5..2586
                     /label=From pC0.026
                     /note="/color=white"
                     /note="/indicates_part=True"
                     /note="/source=pC0.026"
     promoter        5..450
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             451..1872
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders
                     bacterialgrowth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     misc_feature    2587..2590
                     /label=AGGT
                     /note="/color=#f7e8f7"
     misc_feature    2591..3800
                     /label=From pC0.027
                     /note="/color=white"
                     /note="/indicates_part=True"
                     /note="/source=pC0.027"
     CDS             complement(2875..3690)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    3691..3800
                     /label=promoter
     misc_feature    3801..3804
                     /label=GCTT
                     /note="/color=#f7e8f7"
     misc_feature    3805..4682
                     /label=From gblock_OriTOriR
                     /note="/color=white"
                     /note="/indicates_part=True"
                     /note="/source=gblock_OriTOriR"
     misc_feature    4675..4681
                     /label=SapI site
     promoter        4713..4842
                     /label=BBa_J72163 GlpT Promoter
                     /note="label: BBa_J72163 GlpT Promoter"
     primer_bind     4713..4732
                     /label=CG17
     RBS             4843..4878
                     /label=RBS
                     /note="label: RBS"
     RBS             4861..4872
                     /label=RBS
                     /note="note: strong bacterial ribosome binding site
                     (Elowitz and Leibler, 2000)"
     CDS             4879..5577
                     /label=mScarlet
                     /note="codon_start: 1 label: mScarlet transl_table: 1
                     translation:
                     MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFSW
                     DILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGAVTVTQDTSLEDGT
                     LIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKMALRLKDGGRYLAD
                     FKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHSTGGMDELYK*"
     terminator      5578..5706
                     /label=BBa_B0015 Terminator
                     /note="label: BBa_B0015 Terminator"
     terminator      5586..5657
                     /label=rrnB T1 terminator
                     /note="label: rrnB T1 terminator note: pLannotate note:
                     /database=snapgene note: /fragment=False note:
                     /identity=100.0 note: /match_length=100.0 note:
                     /other=terminator"
     terminator      5673..5700
                     /label=T7Te terminator
                     /note="label: T7Te terminator note: pLannotate note:
                     /database=snapgene note: /fragment=False note:
                     /identity=100.0 note: /match_length=100.0 note:
                     /other=terminator"
     primer_bind     complement(5685..5706)
                     /label=CG18
     misc_feature    5734..5747
                     /label=From gblock_OriTOriR
                     /note="/color=white"
                     /note="/indicates_part=True"
                     /note="/source=gblock_OriTOriR"
     misc_feature    5738..5744
                     /label=SapI site
ORIGIN
        1 aatgcacata tacctgccgt tcactattat ttagtgaaat gagatattat gatattttct
       61 gaattgtgat taaaaaggca actttatgcc catgcaacag aaactataaa aaatacagag
      121 aatgaaaaga aacagataga ttttttagtt ctttaggccc gtagtctgca aatcctttta
      181 tgattttcta tcaaacaaaa gaggaaaata gaccagttgc aatccaaacg agagtctaat
      241 agaatgaggt cgaaaagtaa atcgcgcggg tttgttactg ataaagcagg caagacctaa
      301 aatgtgtaaa gggcaaagtg tatactttgg cgtcacccct tacatatttt aggtcttttt
      361 ttattgtgcg taactaactt gccatcttca aacaggaggg ctggaagaag cagaccgcta
      421 acacagtaca taaaaaagga gacatgaacg atgaacatca aaaagtttgc aaaacaagca
      481 acagtattaa cctttactac cgcactgctg gcaggaggcg caactcaagc gtttgcgaaa
      541 gaaacgaacc aaaagccata taaggaaaca tacggcattt cccatattac acgccatgat
      601 atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc aagttcctga attcgattcg
      661 tccacaatta aaaatatctc ttctgcaaaa ggcctggacg tttgggacag ctggccatta
      721 caaaacgctg acggcactgt cgcaaactat cacggctacc acatcgtctt tgcattagcc
      781 ggagatccta aaaatgcgga tgacacatcg atttacatgt tctatcaaaa agtcggcgaa
      841 acttctattg acagctggaa aaacgctggc cgcgtcttta aagacagcga caaattcgat
      901 gcaaatgatt ctatcctaaa agaccaaaca caagaatggt caggttcagc cacatttaca
      961 tctgacggaa aaatccgttt attctacact gatttctccg gtaaacatta cggcaaacaa
     1021 acactgacaa ctgcacaagt taacgtatca gcatcagaca gctctttgaa catcaacggt
     1081 gtagaggatt ataaatcaat ctttgacggt gacggaaaaa cgtatcaaaa tgtacagcag
     1141 ttcatcgatg aaggcaacta cagctcaggc gacaaccata cgctgagaga tcctcactac
     1201 gtagaagata aaggccacaa atacttagta tttgaagcaa acactggaac tgaagatggc
     1261 taccaaggcg aagaatcttt atttaacaaa gcatactatg gcaaaagcac atcattcttc
     1321 cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac gcacggctga gttagcaaac
     1381 ggcgctctcg gtatgattga gctaaacgat gattacacac tgaaaaaagt gatgaaaccg
     1441 ctgattgcat ctaacacagt aacagatgaa attgaacgcg cgaacgtctt taaaatgaac
     1501 ggcaaatggt acctgttcac tgactcccgc ggatcaaaaa tgacgattga cggcattacg
     1561 tctaacgata tttacatgct tggttatgtt tctaattctt taactggccc atacaagccg
     1621 ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc ctaacgatgt aacctttact
     1681 tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg tcgtgattac aagctatatg
     1741 acaaacagag gattctacgc agacaaacaa tcaacgtttg cgccaagctt cctgctgaac
     1801 atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc ttgaacaagg acaattaaca
     1861 gttaacaaat aaaaacgcaa aagaaaatgc cgatatccta ttggcatttt cttttatttc
     1921 ttatcaacat aaaggtgaat cccatatgaa ctatataaaa gcaggcaaat ggctaaccgt
     1981 attcctaacc ttttggtaat gactccaact tattgatagt gttttatgtt cagataatgc
     2041 ccgatgactt tgtcatgcag ctccaccgat tttgagaacg acagcgactt ccgtcccagc
     2101 cgtgccaggt gctgcctcag attcaggtta tgccgctcaa ttcgctgcgt atatcgcttg
     2161 ctgattacgt gcagctttcc cttcaggcgg gattcataca gcggccagcc atccgtcatc
     2221 catatcacca cgtcaaaggg tgacagcagg ctcataagac gccccagcgt cgccatagtg
     2281 cgttcaccga atacgtgcgc aacaaccgtc taccggagac tgtcatacgc gtaaaacagc
     2341 cagcgctggc gcgatttagc cccgacatag ccccactgtt cgtccatttc cgcgcagacg
     2401 atgacgtcac tgcccggctg tatgcgcgag gttaccgact gcggcctgag ttttttaagt
     2461 gacgtaaaat cgtgttgagg ccaacgccca taatgcgggc tgttgcccgg catccaacgc
     2521 cattcatggc catatcaatg attttctggt gcgtaccggg ttgagaagcg gtgtaagtga
     2581 actgcaaggt ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa
     2641 tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg
     2701 tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa
     2761 gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc
     2821 ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa
     2881 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2941 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     3001 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     3061 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     3121 cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt
     3181 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     3241 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     3301 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     3361 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     3421 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     3481 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     3541 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     3601 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     3661 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     3721 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     3781 ttgagacaca acgtggcttt gcttgtacac cttgataggt gggctgccct tcctggttgg
     3841 cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc
     3901 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
     3961 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
     4021 ccaaggaaag tctacacgaa ccctttggca aaatatcaaa ggatcttctt gagatccttt
     4081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggcgctctt
     4681 catgctcgga gagagacctt tgccttgtct tcgaaagtga aacgtgattt catgcgtcat
     4741 tttgaacatt ttgtaaatct tatttaataa tgtgtgcggc aattcacatt taatttatga
     4801 atgttttctt aacatcgcgg caactcaaga aacggcaggt tcggatctta gctactagag
     4861 aaagaggaga aatactagat ggttagcaaa ggcgaggcgg ttatcaagga gtttatgcgt
     4921 tttaaggttc acatggaggg tagcatgaac ggtcacgagt tcgagatcga gggtgaaggc
     4981 gagggtcgtc cgtacgaagg cacccagacc gcgaagctga aagtgaccaa gggtggcccg
     5041 ctgccgttca gctgggacat cctgagcccg cagttcatgt atggcagccg tgcgtttacc
     5101 aaacacccgg cggacattcc ggattactat aagcaaagct tcccggaagg ttttaaatgg
     5161 gagcgtgtta tgaacttcga agatggtggc gcggtgaccg ttacccagga caccagcctg
     5221 gaggatggca ccctgattta caaggtgaaa ctgcgtggca ccaactttcc gccggatggt
     5281 ccggttatgc agaagaaaac gatgggttgg gaagcgagca ccgagcgtct gtatccggaa
     5341 gatggcgtgc tgaagggtga tatcaaaatg gcgctgcgtc tgaaggacgg tggccgttac
     5401 ctggcggatt ttaagaccac ctataaagcg aagaaaccgg tgcaaatgcc gggtgcgtac
     5461 aacgttgacc gtaaactgga tattaccagc cacaacgagg attataccgt ggttgagcaa
     5521 tatgagcgta gcgagggtcg ccacagcacc ggcggcatgg acgaactgta taagtgacca
     5581 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     5641 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     5701 tttatagaag acaagggaaa ggtctctcgc tgagatagaa gagcacg
//
