LOCUS       Exported                4420 bp ds-DNA     circular SYN 16-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4420)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Nov 17, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4420
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      22..858
                     /label=more stringent p15A
                     /label=more\stringent\p15A
     CDS             complement(930..1745)
                     /label=Kanamycin phosphotransferase
                     /label=Kanamycin\phosphotransferase
     terminator      1846..1891
                     /label=L3S3P21
                     /note="Geneious type: terminator"
     misc_feature    1898..1901
                     /label=4bp_scar5
     promoter        1922..2011
                     /label=pCymRC promoter
                     /label=pCymRC
     misc_feature    1922..1953
                     /label=CymO
                     /note="Geneious type: operator"
     promoter        1945..1950
                     /label=-35 promoter
                     /label=-35
     promoter        1968..1973
                     /label=-10 promoter
                     /label=-10
     misc_feature    1980..2011
                     /label=CymO
                     /note="Geneious type: operator"
     misc_feature    2013..2086
                     /label=RiboJ53
                     /note="Geneious type: riboj"
     primer_bind     complement(2052..2086)
                     /label=GML_amplify_Cym
     RBS             2087..2130
                     /note="TIR: 41546"
     primer_bind     2116..2146
                     /label=oGML0592_f
                     /note="amplify CYP067"
     CDS             2131..3330
                     /codon_start=1
                     /product="CYP067"
                     /label=CYP067
                     /db_xref="UniProtKB/Swiss-Prot:Q98A71"
                     /translation="MPAAPTQLDRLSSAILRQGGMARVSLPGDVVTWAAARHQTLRQML
                     SDQRFNKDWRQWRALQDGEIPEDHPLIGICKVDNMTTAHGADHRRLRGLLSSSFAPSRI
                     ALLAPRVEQCVDRLLAEMAQRGGSADLMSEFAAPLPTNVIAELFGLPDEQREEIVALTY
                     SLASTSATAEEVRQTRQRIPEFFRRLIALKRGQLGDDLASALIVARDKGELVSDTELID
                     MLFMVLSAGFVTTAGVIGNGVLALLTHPQQLHLVRSGQVPWSQAIEEILRWGTSAANLP
                     FRYATQDVEIDGCLVRRGDAVLMAFHAANRDEKAFGPGANRFDVTRRHNPHLSFGEGPH
                     SCLGAALARLELRCAFPPLFGRLEDLALTIAAEDVVYMPSYVIRCPQRLPVSFRPSVA*
                     "
     primer_bind     complement(3310..3328)
                     /label=oGML0592_r
                     /note="amplify CYP067"
     terminator      3372..3418
                     /label=L3S3P41
     misc_feature    3414
                     /label=C to G
     misc_feature    3428..3431
                     /label=4bp_scar6
     promoter        3447..3528
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3465..3470
                     /label=-35 promoter
                     /label=-35
     misc_feature    3466
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3488..3493
                     /label=-10 promoter
                     /label=-10
     CDS             3529..4140
                     /label=CymR
     misc_feature    3826..3828
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    3828
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3856..3858
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    3856
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3910..3912
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    3911
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4039..4041
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4039
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4150..4404
                     /label=induction operon terminator region
ORIGIN
        1 actgactcgc tgcacgaggc agacctcagc gctagcggag tgtatactgg cttactatgt
       61 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
      121 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
      181 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
      241 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
      301 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
      361 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
      421 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
      481 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
      541 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      601 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
      661 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
      721 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
      781 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
      841 agaagatcat cttattaagg ggtctgacgc tcagtggaac gaaaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacct tagaaaaact catcgagcat caaatgaaac
      961 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1021 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1081 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1141 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     1201 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1261 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1321 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1381 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1441 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1501 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1561 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1621 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1681 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     1741 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1801 atatttttat cttgtgcaat gtacatcaga gattttgaga cacaaccaat tattgaaggc
     1861 ctccctaacg gggggccttt ttttgtttct ggtctcccaa gcgcttaacg atcgttggct
     1921 gaacaaacag acaatctggt ctgtttgtat tatggaaaat ttttctgtat aatagattca
     1981 acaaacagac aatctggtct gtttgtatta tcagcggtca acgcatgtgc tttgcgttct
     2041 gatgagacag tgatgtcgaa accgcctcta caaataattt tgtttaggag gccccaatta
     2101 aatttaccat tgatataagg agtttttttt atgccggcag cgccaaccca actggatcgt
     2161 ctgagctcag ccattttgcg ccagggtggt atggctcgcg tttcgttgcc aggcgacgtt
     2221 gtcacctggg ccgctgcgcg gcatcagacc ttacgccaga tgctctcaga tcagcgcttt
     2281 aacaaagatt ggcgtcagtg gcgcgccctc caagacggag agattcccga ggatcatcct
     2341 cttattggta tctgtaaagt tgataatatg accaccgcac atggtgctga ccaccgacgc
     2401 ttacgtgggc tcctgtctag cagctttgca cccagtcgta tcgcactgtt agcgccgcgt
     2461 gttgaacagt gtgtggaccg tttactggct gaaatggccc agcgtggtgg ttcagcggac
     2521 ctgatgtctg aattcgccgc tccgctgccc accaatgtga ttgcggagct gtttggcctc
     2581 ccagacgaac agcgcgagga aatcgtagca ctgacctatt ccctggcttc cacgtctgct
     2641 accgcggaag aagtacgcca gacgagacaa cgtattccgg aattcttccg acgtctgatt
     2701 gccctgaagc gtggacagct cggcgacgat ttagcatcag ctcttattgt cgcgcgcgac
     2761 aaaggtgaac tggtttctga tacggaactg atcgatatgc tgtttatggt gttatccgcc
     2821 ggcttcgtca cgaccgctgg cgtgatcggc aacggtgtcc tggcgttgct tacgcacccg
     2881 caacagctgc atttagtgcg atctggtcaa gtgccctggt cgcaggctat cgaggaaatt
     2941 ttacgctggg gtacgagtgc ggctaactta ccgtttcggt acgcgaccca agacgtcgaa
     3001 attgatggtt gtctggttcg ccgtggggat gccgtattga tggcgtttca tgctgctaat
     3061 cgcgatgaaa aagcgttcgg gccgggtgcg aatcgctttg acgtgacccg ccgccataat
     3121 ccgcacttgt cattcggtga aggtccgcat tcctgtttgg gggcggcact tgcaagactg
     3181 gaattgcgat gtgcctttcc gccgttattc gggcgtctgg aagatttagc tcttacgatc
     3241 gctgcagagg acgtggtcta catgccgtct tacgtcattc gctgcccaca gcgcctcccc
     3301 gtgtcttttc ggccctcagt cgcctaataa ggatgagctc tacaaataag cagaggtggt
     3361 tgtgttgcga aaaaaaaaaa aaacacccta acgggtgttt tttttttttt ggtgtcccca
     3421 cgtgtggcgc tggagaccgt ccaatggcgg cgcgccatcg aatggtgcaa aacctttcgc
     3481 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatat gagcccgaaa
     3541 cgtcgtaccc aggcagaacg tgcaatggaa acccagggta aactgattgc agcagcactg
     3601 ggtgttctgc gtgaaaaagg ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc
     3661 ggtgttagcc gtggtgcaca gagccatcat tttccgacca aactggaact gctgctggca
     3721 acctttgaat ggctgtatga gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg
     3781 aaaccggaag atgatgttat tcagcagatg ctggatgatg cagcagattt ttttctggat
     3841 gatgatttta gcatcggcct ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt
     3901 gaaggtattc tgcgtaccgt tgaacgtaat cgttttgttg ttgaagatat gtggctgggt
     3961 gtgctggtga gccgtggtct gagccgtgat gatgccgaag atattctgtg gctgattttt
     4021 aacagcgttc gtggtctgac agttcgtagc ctgtggcaga aagataaaga acgttttgaa
     4081 cgtgtgcgta atagcaccct ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga
     4141 taaggatcct aattggtaac gaatcagaca attgacggct cgagggagta gcatagggtt
     4201 tgcagaatcc ctgcttcgtc catttgacag gcacattatg catcgatgat aagctgtcaa
     4261 acatgagcag atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     4321 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     4381 gctcatgagc aaatatttta tctgaggtgc ttcctcgctc
//
