LOCUS       Exported                4447 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 4447)
  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..4447
                     /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..2124
                     /note="TIR: 48350"
     CDS             2125..3357
                     /codon_start=1
                     /product="CYP069"
                     /label=CYP069
                     /db_xref="UniProtKB/Swiss-Prot:Q06069"
                     /translation="MKEVIAVKEITRFKTRTEEFSPYAWCKRMLENDPVSYHEGTDTWN
                     VFKYEDVKRVLSDYKHFSSVRKRTTISVGTDSEEGSVPEKIQITESDPPDHRKRRSLLA
                     AAFTPRSLQNWEPRIQEIADELIGQMDGGTEIDIVASLASPLPIIVMADLMGVPSKDRL
                     LFKKWVDTLFLPFDREKQEEVDKLKQVAAKEYYQYLYPIVVQKRLNPADDIISDLLKSE
                     VDGEMFTDDEVVRTTMLILGAGVETTSHLLANSFYSLLYDDKEVYQELHENLDLVPQAV
                     EEMLRFRFNLIKLDRTVKEDNDLLGVELKEGDSVVVWMSAANMDEEMFEDPFTLNIHRP
                     NNKKHLTFGNGPHFCLGAPLARLEAKIALTAFLKKFKHIEAVPSFQLEENLTDSATGQT
                     LTSLPLKASRM"
     terminator      3399..3445
                     /label=L3S3P41
     misc_feature    3441
                     /label=C to G
     misc_feature    3455..3458
                     /label=4bp_scar6
     promoter        3474..3555
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3492..3497
                     /label=-35 promoter
                     /label=-35
     misc_feature    3493
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3515..3520
                     /label=-10 promoter
                     /label=-10
     CDS             3556..4167
                     /label=CymR
     misc_feature    3853..3855
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    3855
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3883..3885
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    3883
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3937..3939
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    3938
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4066..4068
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4066
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4177..4431
                     /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 tgtttagtag ggacgtataa
     2101 cccaaatccg gcgggagaga ttttatgaag gaagtgatcg cggtgaaaga aataacccgt
     2161 tttaaaacgc gtactgaaga attttcgccc tacgcttggt gcaaacgtat gcttgaaaac
     2221 gaccctgtta gctatcatga aggcacggat acttggaacg tgtttaaata tgaggatgtg
     2281 aaacgcgtac tgagtgacta taaacatttt agctctgtta gaaaacgtac taccatcagt
     2341 gtcgggacgg actcggaaga aggttccgtg ccagagaaaa tccagattac ggaaagcgac
     2401 ccaccggatc atcgcaaacg ccggagtctg ttggccgcgg ccttcacccc cagatctctg
     2461 cagaactggg aacctcgcat ccaagaaatc gccgatgaac tgattggtca gatggatggc
     2521 ggaacagaga ttgatatagt ggccagcctg gccagccctt taccgattat cgtaatggcc
     2581 gacctgatgg gcgtgccgag caaagaccgc ctgctgttta agaagtgggt ggataccctg
     2641 tttctgccgt ttgatcgcga gaaacaagaa gaggtggata aactgaagca agtcgcggcc
     2701 aaagaatatt atcagtatct ctatccgatc gtggttcaaa agcgactgaa ccctgcagac
     2761 gacatcattt ctgacttgct gaaaagcgaa gttgatggag agatgttcac cgatgacgag
     2821 gtggtgcgca ctacgatgct tatcttaggt gctggtgtgg aaactacgtc acacctcctg
     2881 gcgaacagct tttatagcct gctgtacgac gataaagaag tttaccaaga attgcatgag
     2941 aacttagatc tggtcccgca ggcggtcgag gaaatgttac gcttccgttt caacttgatt
     3001 aagctggacc gtacggtcaa agaagataat gatctcctgg gcgttgagct gaaagaaggt
     3061 gattccgtcg tcgtctggat gagcgccgcg aatatggatg aggagatgtt cgaggacccg
     3121 tttactctga atatccatcg acctaacaac aagaaacacc tgacctttgg caatggtccg
     3181 catttctgtc tgggtgcgcc cttagcaaga ctcgaagcca aaatcgccct gaccgcgttc
     3241 ctcaagaaat ttaaacatat tgaagcagtg cctagctttc agctggaaga gaatttaacc
     3301 gactcggcta cgggccagac gctgacctca ctgcctctga aagcaagtcg tatgtgagga
     3361 tgagctctac aaataagcag aggtggttgt gttgcgaaaa aaaaaaaaaa caccctaacg
     3421 ggtgtttttt tttttttggt gtccccacgt gtggcgctgg agaccgtcca atggcggcgc
     3481 gccatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     3541 ttcagggtgg tgaatatgag cccgaaacgt cgtacccagg cagaacgtgc aatggaaacc
     3601 cagggtaaac tgattgcagc agcactgggt gttctgcgtg aaaaaggtta tgcaggtttt
     3661 cgtattgcag atgttccggg tgcagccggt gttagccgtg gtgcacagag ccatcatttt
     3721 ccgaccaaac tggaactgct gctggcaacc tttgaatggc tgtatgagca gattaccgaa
     3781 cgtagccgtg cacgtctggc aaaactgaaa ccggaagatg atgttattca gcagatgctg
     3841 gatgatgcag cagatttttt tctggatgat gattttagca tcggcctgga tctgattgtt
     3901 gcagcagatc gtgatccggc actgcgtgaa ggtattctgc gtaccgttga acgtaatcgt
     3961 tttgttgttg aagatatgtg gctgggtgtg ctggtgagcc gtggtctgag ccgtgatgat
     4021 gccgaagata ttctgtggct gatttttaac agcgttcgtg gtctgacagt tcgtagcctg
     4081 tggcagaaag ataaagaacg ttttgaacgt gtgcgtaata gcaccctgga aattgcacgt
     4141 gaacgttatg caaaattcaa acgttgataa ggatcctaat tggtaacgaa tcagacaatt
     4201 gacggctcga gggagtagca tagggtttgc agaatccctg cttcgtccat ttgacaggca
     4261 cattatgcat cgatgataag ctgtcaaaca tgagcagatc ctctacgccg gacgcatcgt
     4321 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     4381 tggggaagat cgggctcgcc acttcgggct catgagcaaa tattttatct gaggtgcttc
     4441 ctcgctc
//
