LOCUS       Exported                6328 bp DNA     circular SYN 07-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    T1_entry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6328)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 7, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1330
                     /mol_type="other DNA"
                     /organism="pKDsgRNA-ack"
     source          1330
                     /mol_type="other DNA"
                     /organism="pTKIP-cat"
     source          4508
                     /mol_type="other DNA"
                     /organism="pKDsgRNA-ack"
     source          4508
                     /mol_type="other DNA"
                     /organism="pTKIP-cat"
     promoter        12..65
                     /note="tet promoter"
     promoter        119..242
                     /note="P lac prom"
     promoter        211
                     /note="P"
     CDS             249..572
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="lacZ-alpha"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    573..1219
                     /label=ccdB Kassette aus pDONR201
                     /note="ccdB Kassette aus pDONR201"
     misc_feature    914..1219
                     /label=ccdB
                     /note="ccdB"
     misc_feature    1327..1391
                     /label=gRNA backbone
                     /note="gRNA backbone"
     terminator      1420..1458
                     /note="Terminator (BBa_B1006)"
     CDS             complement(1462..4482)
                     /codon_start=1
                     /label=vioB (sites removed)
                     /note="vioB"
                     /translation="MSLLDFPRLHFRGFARANVPTGNRNTHGNIDIATNAVSMAGEAVD
                     LSRPPAEFHAHLKQLAPRFNAQGKPDPDGIFSQATGYNFCGNNHFSWENARITGVQLRD
                     GEVDTQDALVGAKLGLWGHYNEYLRTTFNRARWIDNNPAQPDTTLIYAGQFTLSDKLAT
                     PNTPTLFTADIAQAHSVRWLGSGHITERSGHFLDEEFGRSRLFQFSVAKQDPHFLFNPD
                     LPLPASMHALQQALADDEVLGLTVQYCLFNMSTPQKPDSPVFYDLAGSIGLWRRGELAT
                     YPAGRLLQPRQGSLGPVLVKVHADRVSFNMPTAIPFTTRDAGAVSEQHPTHALGGKQAL
                     GDLLLHDGAGTVLARIPEQLYRDYWRHHGVFDVPLQHAGAAPGSLSLGSAQAQWDEADW
                     VLQSDSNQLYLEAPNRNKHEQFPQTITVQSRFRGELAAPPSLAEAEDGVLLAVEQQPSP
                     LGHGYTTLTLTGRKPGATRIVLGTGKAKQYLGVRVLPDDWDLDDVPAEQVDYAFLYRHV
                     MSYYELVYPFMSDKVFSLADQCKCETYSRLMWQMCDPQNRDKSYYMPSTRELSLPKSRL
                     FLKYLTQVEAAAAAKAAAPEPAAPHAIGGKAELIDELKKAIDLELSLMLQYLYAAYSIP
                     NYAQGAALVQSGRWLPAELELACGAEDRRRNSGTRGALLEIAHEEMIHYLLVNNVLMAL
                     GEPFYSGTPLLGQQARQRFGLDTEFAFEPFSEHVLARFVRFEWPDYIPTPGKSIATFYI
                     AIRQALAELPGLFESGGGKRGGEHHLFLKELTNRAYPGYQLEVSDRDSALFAIDFVTEQ
                     GEGVAVDSPHFASSHFQRLRAIAGRFSACDKPFEPALPALKNPVLEARADCSVVTDQKA
                     RALMQLYQGCYELTFLMMAHHFAQQPLGSLRRSRLMNASIDIMTGLLRPLSAALMNMPS
                     GLPGRHAGPPVPEPVSSRVSSDYSLGCDMLAQRCLALAQYARSLESDAIGMAPIEMLEF
                     FNQQLTDLSRGKMSREA"
     misc_feature    complement(4484..4488)
                     /label=spacer
                     /note="spacer"
     RBS             4489..4494
                     /label=RBS
     misc_feature    complement(4495..4504)
                     /label=enhancer_sequence
                     /note="enhancer_sequence"
     CDS             4720..5511
                     /codon_start=1
                     /label=NPTII
                     /note="NPTII"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    5539..6328
                     /label=pUC ori
                     /note="pUC ori"
ORIGIN
        1 gcagaagacc atccctatca gtgatagaga ttgacatccc tatcagtgat agagatactg
       61 agcactgaga cccagtggta tgggcagctg gcacgacagg tttcccgact ggaaagcggg
      121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      241 aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc
      301 cccgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      361 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      421 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      481 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      541 ctctcagtac aatctgctct gatgccgcat agtcgcgtgg atccggctta ctaaaagcca
      601 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt
      661 atacccgaag tatgtcaaaa agaggtgtgc tatgaagcag cgtattacag tgacagttga
      721 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc
      781 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag
      841 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac
      901 agggactggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
      961 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
     1021 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
     1081 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgcctg tctccgttat
     1141 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
     1201 gatgttctgg ggaatataat taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     1261 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccc actcaagtgg
     1321 gtctcagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     1381 aaaagtggca ccgagtcggt gcttttttta cggtcatgta aaaaaaaacc ccgcccctga
     1441 cagggcgggg ttttttttca gtcaagcctc tcttgacatc tttccccgag ataaatcggt
     1501 aagttgctga ttaaaaaact ccaacatttc tatcggcgcc atgccgatag catcgctctc
     1561 caggctgcgc gcgtactgcg ccagcgccag gcatctctgc gccagcatgt cgcagcccag
     1621 gctgtagtcg ctgctgaccc ggctgctgac cggctcgggc acgggcggtc cagcatggcg
     1681 gccaggcagg ccggacggca tgttcatcag ggccgccgac agggggcgca acaggcctgt
     1741 catgatgtcg atggacgcgt tcatcaggcg cgagcggcgc aggctgccca gcggctgctg
     1801 cgcaaaatgg tgcgccatca tcaggaaggt cagttcatag cagccctgat acagctgcat
     1861 cagcgcgcgc gccttctgat cggtcaccac gctgcagtcc gcgcgcgctt ccagcacggg
     1921 attcttcagc gccggcagcg ccggttcgaa cggcttgtcg caggccgaaa acctgccggc
     1981 gatggcgcgc agccgctgga agtgcgagga ggcgaaatgc ggcgaatcga cggccacgcc
     2041 ttcgccctgt tccgtgacga aatcgatggc gaacaacgcg ctgtcgcggt cggatacttc
     2101 cagctggtag ccgggatagg cgcggttggt cagttctttc aggaacaggt ggtgctcgcc
     2161 gccgcgcttg ccgccgccgc tttcgaacag gccgggcagc tcggccaggg cctggcggat
     2221 cgcgatatag aaggtggcga tggatttgcc cggcgtggga atgtagtcgg gccattcaaa
     2281 acgcacgaag cgggccagca cgtgttcgga aaatggttcg aacgcaaatt ccgtgtccag
     2341 gccgaaacgc tggcgcgcct gctggcccag cagcggggta ccgctgtaaa acggttcgcc
     2401 aagcgccatc aatacattgt tcaccaataa gtagtgaatc atttcttcat gggcgatttc
     2461 cagcagcgcg ccgcgcgtgc cgctgttgcg gcgccgatcc tcggcgccac acgctaactc
     2521 cagttccgcg ggaagccagc gaccagactg caccaacgcg gcaccttgcg cataattcgg
     2581 gatgctatac gcagcgtaca gatactgcag catcaggctt aattccaggt caatcgcctt
     2641 cttcagctca tctatcagtt cagctttgcc gccaatcgca tgaggcgccg ctggttccgg
     2701 agccgccgct tttgcggccg cggcggcttc tacttgggtc agatatttca gaaacaagcg
     2761 ggatttaggc agcgacagtt cgcgagtgct gggcatgtag tagcttttgt cccgattttg
     2821 cgggtcacac atctgccaca tcaggcgtga atacgtttcg catttgcact gatcagccag
     2881 gctgaacacc ttatcggaca taaaggggta aaccaattcg tagtaactca tcacatggcg
     2941 gtacaagaag gcatagtcca cttgttctgc tggaacgtcg tctaagtccc agtcatccgg
     3001 cagaacccgc acgccaagat actgtttcgc tttgccagtt ccaaggacaa tgcgcgttgc
     3061 tcccggttta cgacctgtga gggtcagcgt ggtatagccg tggcctaacg gactcggttg
     3121 ctgctcgacg gctaagagga cgccatcttc cgcttcggcc agtgacggag gtgcagccag
     3181 ttcaccacgg aaacgcgatt gaaccgtgat ggtctgcgga aactgctcgt gtttgttgcg
     3241 attcggtgcc tccaggtaca gctggttaga atcgctttgg aggacccaat cggcttcatc
     3301 ccactgtgcc tgggctgaac cgagcgaaag agaacctggt gcagctcccg catgctgtaa
     3361 tggaacatca aacacgccat gatgacgcca atagtcgcga tacaactgtt ccggaatacg
     3421 ggcgagtacg gtccctgccc catcgtgcag cagcagatcg cccaatgctt gtttaccacc
     3481 cagcgcatgt gtcggatgct gctcggatac agccccggca tcacgggtag taaacgggat
     3541 tgcggtaggc atgttgaaac tcacgcgatc cgcatgcact ttcaccagca ccggccccag
     3601 gctgccctgg cgcggctgca gcaggcggcc ggccggatag gtggccagct cgccgcgccg
     3661 ccacaggccg atgctgccgg ccaggtcgta gaacacgggc gaatcgggtt tttgcggcgt
     3721 cgacatattg aacaggcagt attgcaccgt caggcccagc acctcgtcgt cggccagggc
     3781 ttgctgcaag gcatgcatac tggccggcag cggcaggtcc ggattgaaca ggaaatgcgg
     3841 gtcctgcttg gccacggaaa actggaacag cctggagcgg ccgaattcct cgtccaggaa
     3901 atgcccgctg cgttccgtga tgtggccgct gccgagccag cgtaccgagt gcgcctgcgc
     3961 gatgtcggcc gtgaacagcg tgggcgtatt gggcgtggcc agcttgtcgc tcaaggtgaa
     4021 ctggcccgcg tagatcagcg tggtgtcggg ctgcgccggg ttgttgtcga tccaccttgc
     4081 gcggttgaac gtcgtgcgca ggtactcgtt gtagtggccc cacaggccca gcttggcgcc
     4141 caccagcgcg tcctgggtat cgacctcgcc atcacgcaac tggacgcccg tgatccgcgc
     4201 gttttcccac gagaaatggt tgttcccgca aaaattatag cctgtcgcct ggctgaagat
     4261 gccgtccgga tcgggcttgc cctgtgcgtt gaagcggggg gcgagctgtt tcaggtgcgc
     4321 atggaattcg gctggcggcc ggctcaggtc gacagcctcg cccgccatcg ataccgcatt
     4381 cgtggcgata tcgatgttgc cgtgcgtatt gcgattcccc gtcggcacat tggcgcgggc
     4441 aaaaccccga aaatgcaggc gggggaagtc aagtaggctc attacgactc tcctctgtaa
     4501 aagcgtcctt gtcttccgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4561 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tggctaaaat gagaatatca
     4741 ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct
     4801 cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac
     4861 agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg
     4921 gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat
     4981 ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc
     5041 cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata
     5101 tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg
     5161 aataacgatc tggccgatgt ggattgcgaa aactgggaag aggacactcc atttaaagat
     5221 ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc
     5281 cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt
     5341 gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg
     5401 atcagggagg atatcgggga agaacagtat gtcgagctat tttttgactt actggggatc
     5461 aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt ttagctgtca
     5521 gaccaagttt actccggggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5581 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5641 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5701 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5761 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5821 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5881 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5941 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6001 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6061 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6121 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6181 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6241 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6301 taaagtatat atgagtaaac ttggtctg
//
