LOCUS       Exported                4561 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 4561)
  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..4561
                     /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..2115
                     /note="TIR: 55061"
     primer_bind     2112..2133
                     /label=oGML0594_f
                     /note="amplify CYP073"
     CDS             2116..3471
                     /codon_start=1
                     /product="CYP073"
                     /label=CYP073
                     /db_xref="UniProtKB/Swiss-Prot:B5GRC6"
                     /translation="MTGGTIARVPGALPLIGHTGAFLKSPLEFLKGLSAHGELVELRFG
                     RKRVLVVCDPDLTQEMLRRDRVYDKGGPIYDRMREAAGNGLATCPSRDHRRQRRTIQPA
                     FHHSRLPGYARTMTEVIDEAVRGWDGPEVDLCHETRSIASSVLLTTLFGTAFAPEARDA
                     LARDLETILRGGLRRAVLPEALSALPTPANLRYQRARRRARGMMADLIRAVRAGDGSGD
                     SLFATLVNQHGAGTGDLPMTDEELVDQAITMYVAGTETTAGVVCWALHLAAQHPQAGRR
                     LADECRSVLGHRAATWEDLPHLPYTRQTLLEALRLNPPGWLITRLTTTDDARLGDHAVP
                     RGSTIVYSAYLLSRLPARYEEPEHFAPERWAPGTMGAKVPDGVFGGGARKCIGDEYALI
                     EGVLMLATITARWDVRTAPDAPKRPHLPALTLNPTRLMATVTPAAAPDGSR*"
     primer_bind     complement(3451..3469)
                     /label=oGML0594_r
                     /note="amplify CYP073"
     terminator      3513..3559
                     /label=L3S3P41
     misc_feature    3555
                     /label=C to G
     misc_feature    3569..3572
                     /label=4bp_scar6
     promoter        3588..3669
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3606..3611
                     /label=-35 promoter
                     /label=-35
     misc_feature    3607
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3629..3634
                     /label=-10 promoter
                     /label=-10
     CDS             3670..4281
                     /label=CymR
     misc_feature    3967..3969
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    3969
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3997..3999
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    3997
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4051..4053
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    4052
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4180..4182
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4180
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4291..4545
                     /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 tgtttaggca ctgtacacat
     2101 aaggaggtat cttttatgac cggcggcacc atcgcgcgcg ttccgggagc attgccgtta
     2161 ataggtcaca cgggcgcgtt cctgaaatcc ccgctggaat ttctgaaagg tctgtccgcg
     2221 catggcgaat tggttgaact gcgcttcggc agaaaacgag ttttagttgt atgcgaccct
     2281 gaccttacgc aggagatgtt acgccgtgat cgggtgtatg ataaaggggg ccccatttac
     2341 gatcgcatgc gcgaggcggc cggaaatggc ttggcaactt gcccgagtcg ggaccaccga
     2401 cggcagcgcc ggactatcca accggcgttt catcatagtc gcctgccggg ttatgcgcgc
     2461 accatgactg aagtaatcga tgaggcagtt cgcggttggg acggcccgga ggtagacctc
     2521 tgtcacgaaa cgcgcagcat tgcgagcagc gtcctgctga caactctgtt tggtaccgcg
     2581 tttgctccgg aggcccgtga cgcgttagcg cgcgatctgg aaaccatcct gcgaggcggc
     2641 cttcgccgcg ccgtactgcc tgaagctctg tcggccctgc ctacccccgc gaacttacgt
     2701 tatcagcgtg cgcgccgccg tgccagaggt atgatggctg acctcatccg tgcagtacgc
     2761 gcaggcgacg gatctggtga ttcactgttc gccaccctgg taaatcaaca cggcgcggga
     2821 acgggcgacc tgccaatgac agatgaggaa ttagttgatc aggctattac catgtacgtc
     2881 gcgggtacgg aaacaaccgc aggagtggta tgttgggcac tgcatctggc agcacagcat
     2941 ccgcaagccg gccgtcgcct ggctgatgaa tgccgtagcg tgctcggtca tcgcgcggca
     3001 acctgggagg atcttcctca tctgccgtac actcgtcaga cgctcttaga agcgctgcgc
     3061 ctcaaccctc cagggtggct tatcacccgg ctgaccacga cggatgatgc acgattaggc
     3121 gaccatgcgg taccacgggg tagtaccatc gtctacagcg catacctgct ctcccggctg
     3181 ccggcgcgat acgaagaacc tgaacatttt gcaccggagc gttgggcgcc gggtaccatg
     3241 ggtgcaaaag ttccagacgg agtattcggt ggtggtgcgc gtaaatgcat cggagatgaa
     3301 tatgcgctga ttgaaggtgt cctgatgctc gcaacaatca ccgcccgttg ggacgttcgt
     3361 actgccccgg atgctccaaa gcgcccgcat ctgcccgcgc ttacgctgaa tccgacccgt
     3421 ttgatggcga ctgttacccc agcagcggcg ccagatggct cgcgctaata aggatgagct
     3481 ctacaaataa gcagaggtgg ttgtgttgcg aaaaaaaaaa aaaacaccct aacgggtgtt
     3541 tttttttttt tggtgtcccc acgtgtggcg ctggagaccg tccaatggcg gcgcgccatc
     3601 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     3661 gtggtgaata tgagcccgaa acgtcgtacc caggcagaac gtgcaatgga aacccagggt
     3721 aaactgattg cagcagcact gggtgttctg cgtgaaaaag gttatgcagg ttttcgtatt
     3781 gcagatgttc cgggtgcagc cggtgttagc cgtggtgcac agagccatca ttttccgacc
     3841 aaactggaac tgctgctggc aacctttgaa tggctgtatg agcagattac cgaacgtagc
     3901 cgtgcacgtc tggcaaaact gaaaccggaa gatgatgtta ttcagcagat gctggatgat
     3961 gcagcagatt tttttctgga tgatgatttt agcatcggcc tggatctgat tgttgcagca
     4021 gatcgtgatc cggcactgcg tgaaggtatt ctgcgtaccg ttgaacgtaa tcgttttgtt
     4081 gttgaagata tgtggctggg tgtgctggtg agccgtggtc tgagccgtga tgatgccgaa
     4141 gatattctgt ggctgatttt taacagcgtt cgtggtctga cagttcgtag cctgtggcag
     4201 aaagataaag aacgttttga acgtgtgcgt aatagcaccc tggaaattgc acgtgaacgt
     4261 tatgcaaaat tcaaacgttg ataaggatcc taattggtaa cgaatcagac aattgacggc
     4321 tcgagggagt agcatagggt ttgcagaatc cctgcttcgt ccatttgaca ggcacattat
     4381 gcatcgatga taagctgtca aacatgagca gatcctctac gccggacgca tcgtggccgg
     4441 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
     4501 agatcgggct cgccacttcg ggctcatgag caaatatttt atctgaggtg cttcctcgct
     4561 c
//
