LOCUS       Exported                4659 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 4659)
  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..4659
                     /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"
     RBS             2087..2116
                     /label=RBS
                     /note="strength = 42839"
     CDS             2117..3568
                     /codon_start=1
                     /product="CYP027"
                     /label=CYP027
                     /db_xref="UniProtKB/Swiss-Prot:A0A2M9LE08"
                     /db_xref="Refseq:EMBL-CDS:PJN29587.1"
                     /translation="MKGNLLAMSTATSPSVTGSVTTPRTATTPGRTPGLAPGGVPVLGH
                     LPMILRSRFEFIETVRNAGPVTRVKLGPKTAYFVNDYELLAQILVSDADKFVRGIHFKK
                     MRNMVGNGVVTTSGDLHRRQRRIMLPSFSQRRLAMHLPVMRKIMSEFVAAVPEQQPYDL
                     MGPVMGVGCDIVTSTMLGEKTPPKVLRLVREAVPVFVENAAIQAVDVTGIYKHLPTKSN
                     RDFERLLNAFNEYMYSVIDDKFRNGAGEEAGLLDMLINATDPETGEKFDRTEVRDQAAT
                     ILLASTETTANTISWACYELARHPRIFAECRAEIDALVKDRNWLDIEIGRHDLPALKRV
                     LFEALRMYPSSYLLSRQASVDTTLGGYAIPKDATILYSHYGQQRDERNFPHGDEFDPDR
                     WLDKDGTEVTASAFMPFGFGAYRCLGESVAVLEATYCLAMMVHQWDFALSDYSEPKMNA
                     TITLSPKDLEFLFTKRAESGARDE*"
     primer_bind     2117..2142
                     /label=oGML0587_f
                     /note="amplify CYP027"
     primer_bind     2586..2600
                     /label=seq1_CYP27
     primer_bind     3303..3319
                     /label=seq2_CYP27
     primer_bind     complement(3546..3566)
                     /label=oGML0587_r
                     /note="amplify CYP027"
     terminator      3611..3657
                     /label=L3S3P41
     misc_feature    3653
                     /label=C to G
     misc_feature    3667..3670
                     /label=4bp_scar6
     promoter        3686..3767
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3704..3709
                     /label=-35 promoter
                     /label=-35
     misc_feature    3705
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3727..3732
                     /label=-10 promoter
                     /label=-10
     CDS             3768..4379
                     /label=CymR
     misc_feature    4065..4067
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    4067
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4095..4097
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    4095
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4149..4151
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    4150
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4278..4280
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4278
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4389..4643
                     /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 tgtttacaac taatattaac
     2101 ttaaggaatt attcaaatga aaggaaattt acttgccatg tcaaccgcta ccagccccag
     2161 cgtaaccggc tcggtgacga cccctcgtac ggccaccaca ccgggaagaa ccccgggact
     2221 ggcaccgggt ggcgtacccg tgctgggaca tttaccaatg atcctccgta gccgttttga
     2281 attcatcgaa accgtacgca acgcaggccc tgtgacccgc gttaaactgg gtcccaaaac
     2341 tgcttacttc gttaacgact acgagctgct cgcccaaatc ttagtttctg acgcagacaa
     2401 gtttgtccgt ggcattcact ttaaaaagat gcgcaacatg gtaggaaatg gggtcgtcac
     2461 gactagcggg gatctgcatc gcagacagcg gcgaattatg ctcccgtcct tcagtcagcg
     2521 gcgcctggct atgcaccttc cggtcatgcg taaaattatg agcgaatttg tggccgccgt
     2581 tccggaacag cagccatacg acttgatggg accggtcatg ggtgtggggt gcgacatcgt
     2641 gacttctact atgctgggag agaaaacgcc tccaaaagtc cttcgtctgg tacgtgaggc
     2701 agttcctgtt tttgtggaga acgcggccat tcaagcagtg gacgtcacgg gaatctacaa
     2761 gcatttgccg accaaatcga atcgcgactt cgaacggctc ctgaatgcgt tcaacgagta
     2821 tatgtattca gtgatcgacg acaaattccg caatggcgct ggggaggaag ctggcctctt
     2881 ggatatgtta atcaatgcaa ccgacccaga aacgggtgaa aagtttgatc gcacggaggt
     2941 acgtgaccag gccgcgacca tcctgcttgc gtcaacagaa accaccgcaa acacaatttc
     3001 atgggcgtgc tacgaactgg caagacaccc aagaatcttt gctgagtgtc gggccgaaat
     3061 cgatgctctg gttaaagatc gcaactggtt ggacatcgag atcggtcgtc acgatctgcc
     3121 ggcactcaaa cgcgtacttt ttgaagcgct gcgcatgtat ccgtcctcct atttattaag
     3181 ccgtcaagct agcgtggaca ccactctcgg tggttatgcg attccgaaag atgcgaccat
     3241 tttgtattcg cattatggtc aacagcgcga tgaacgtaat ttcccacacg gcgatgaatt
     3301 cgatcctgat cggtggctgg acaaggatgg tacggaagtg accgcatcgg catttatgcc
     3361 tttcggtttt ggcgcctatc gttgcttggg cgaaagcgtg gcggtgctgg aggccaccta
     3421 ctgtttagcc atgatggtgc accagtggga ttttgcactg tctgactact ccgaaccgaa
     3481 aatgaatgcg acgattaccc tgtcaccgaa agacctggaa tttttattca ccaaacgagc
     3541 tgaatcaggt gcacgcgacg aataataagg atgagctcta caaataagca gaggtggttg
     3601 tgttgcgaaa aaaaaaaaaa aacaccctaa cgggtgtttt tttttttttg gtgtccccac
     3661 gtgtggcgct ggagaccgtc caatggcggc gcgccatcga atggtgcaaa acctttcgcg
     3721 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatatg agcccgaaac
     3781 gtcgtaccca ggcagaacgt gcaatggaaa cccagggtaa actgattgca gcagcactgg
     3841 gtgttctgcg tgaaaaaggt tatgcaggtt ttcgtattgc agatgttccg ggtgcagccg
     3901 gtgttagccg tggtgcacag agccatcatt ttccgaccaa actggaactg ctgctggcaa
     3961 cctttgaatg gctgtatgag cagattaccg aacgtagccg tgcacgtctg gcaaaactga
     4021 aaccggaaga tgatgttatt cagcagatgc tggatgatgc agcagatttt tttctggatg
     4081 atgattttag catcggcctg gatctgattg ttgcagcaga tcgtgatccg gcactgcgtg
     4141 aaggtattct gcgtaccgtt gaacgtaatc gttttgttgt tgaagatatg tggctgggtg
     4201 tgctggtgag ccgtggtctg agccgtgatg atgccgaaga tattctgtgg ctgattttta
     4261 acagcgttcg tggtctgaca gttcgtagcc tgtggcagaa agataaagaa cgttttgaac
     4321 gtgtgcgtaa tagcaccctg gaaattgcac gtgaacgtta tgcaaaattc aaacgttgat
     4381 aaggatccta attggtaacg aatcagacaa ttgacggctc gagggagtag catagggttt
     4441 gcagaatccc tgcttcgtcc atttgacagg cacattatgc atcgatgata agctgtcaaa
     4501 catgagcaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     4561 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     4621 ctcatgagca aatattttat ctgaggtgct tcctcgctc
//
