LOCUS       Exported                4460 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 4460)
  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..4460
                     /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..2116
                     /note="TIR: 25941"
     CDS             2117..3370
                     /codon_start=1
                     /product="CYP071"
                     /label=CYP071
                     /db_xref="UniProtKB/Swiss-Prot:Q9AJE5 "
                     /translation="MADLETKFPRYELISSGKYVDRIPELHELREKSPIAWVPVMDAAF
                     LTRHADIVRVLKDHRMAPANLTQGIRLLSPEQQEELEPLSSAVKKWMGHTVPADHQRFI
                     GLLKRYFTPAMIDRMRPRVRQLSHELLDAVEPAGRMDIVSDIAYPLPACVIAEMLGVPM
                     DNRAQLLAWSADIGAIAEIVSYDRLMECQRSLLAMQDFVLEVVKERRAEPKDDLISMFV
                     AAEREGLVSEAEILSNCVMLLFSGHETTGGLITSGLVQLFDHPDQLELLKSDPDLMPGA
                     VEEMLRLAGPASVISRVSTEPVEVAGSSIRSGAAVPPGADGGKRDPRVFEDPDRLDATR
                     RPNDHLAFATGMFYCLGAALARMEADEFFRILLDRFPDVNPGYETPDWQPVLLISRRLK
                     TLPVNLRGVGGSGAGDE*"
     terminator      3412..3458
                     /label=L3S3P41
     misc_feature    3454
                     /label=C to G
     misc_feature    3468..3471
                     /label=4bp_scar6
     promoter        3487..3568
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3505..3510
                     /label=-35 promoter
                     /label=-35
     misc_feature    3506
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3528..3533
                     /label=-10 promoter
                     /label=-10
     CDS             3569..4180
                     /label=CymR
     misc_feature    3866..3868
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    3868
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3896..3898
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    3896
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3950..3952
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    3951
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4079..4081
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4079
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4190..4444
                     /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 tgtttaagag cttcaataag
     2101 ttaaggagag ttttatatgg cggatctgga aacaaaattt ccgcgttacg aacttattag
     2161 ttctggcaag tatgtggacc gtatcccgga attacacgag ctgcgagaga agagtccgat
     2221 tgcatgggta ccggttatgg acgctgcgtt cctcacgcgc cacgcggata tcgttcgcgt
     2281 tttgaaagat caccgcatgg cccccgctaa cctgacccaa gggattcggc tgctgagtcc
     2341 ggaacagcag gaagaattgg aacccctgag ttcagctgtg aagaaatgga tgggacatac
     2401 cgttcctgcc gaccatcagc gcttcattgg tctgcttaaa cgctatttta ccccggcgat
     2461 gattgaccgt atgcgtccgc gtgttcgaca gctcagtcat gagctcctgg atgcagttga
     2521 accggcaggc cgcatggata tagtatcaga tatcgcgtat ccgctgccgg cgtgcgtgat
     2581 cgctgaaatg ttaggagttc cgatggataa ccgggctcag ttgctggcct ggagcgcgga
     2641 catcggcgcc attgcggaga tcgtcagcta cgaccgcctg atggaatgcc agcgtagtct
     2701 gctggccatg caggactttg tgctcgaagt tgtgaaagaa cgtcgtgccg aaccgaaaga
     2761 tgatctgatc tcaatgttcg tcgcggcgga gcgtgagggc ctcgtcagtg aagccgaaat
     2821 tctgtcgaat tgtgtcatgt tgttgtttag cggacatgag acaaccggag gtttgatcac
     2881 cagtggctta gttcagctgt ttgatcatcc tgatcaactg gaactgctga aatctgatcc
     2941 tgatttgatg ccaggcgctg tcgaggaaat gctgcgtctg gccggtcctg cgagcgttat
     3001 ttcacgtgtt agtactgaac ctgtggaagt agcaggctca agcatccgtt ctggcgctgc
     3061 tgtaccgcct ggtgcggatg gtggtaaacg tgatcctcgt gtctttgagg accccgatcg
     3121 tctggacgct acccgccgcc cgaacgatca cttggcgttt gcgaccggta tgttttactg
     3181 ccttggggcc gctcttgcac gtatggaagc cgatgaattt ttccgcatcc ttctggatcg
     3241 tttcccggat gtgaatccgg gctatgaaac accggactgg cagcctgtgc tgttgatttc
     3301 tcggcggtta aaaaccttac cggtgaattt acgaggggtg ggcggctctg gggcgggaga
     3361 tgagtaataa ggatgagctc tacaaataag cagaggtggt tgtgttgcga aaaaaaaaaa
     3421 aaacacccta acgggtgttt tttttttttt ggtgtcccca cgtgtggcgc tggagaccgt
     3481 ccaatggcgg cgcgccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
     3541 ggaagagagt caattcaggg tggtgaatat gagcccgaaa cgtcgtaccc aggcagaacg
     3601 tgcaatggaa acccagggta aactgattgc agcagcactg ggtgttctgc gtgaaaaagg
     3661 ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc ggtgttagcc gtggtgcaca
     3721 gagccatcat tttccgacca aactggaact gctgctggca acctttgaat ggctgtatga
     3781 gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg aaaccggaag atgatgttat
     3841 tcagcagatg ctggatgatg cagcagattt ttttctggat gatgatttta gcatcggcct
     3901 ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt gaaggtattc tgcgtaccgt
     3961 tgaacgtaat cgttttgttg ttgaagatat gtggctgggt gtgctggtga gccgtggtct
     4021 gagccgtgat gatgccgaag atattctgtg gctgattttt aacagcgttc gtggtctgac
     4081 agttcgtagc ctgtggcaga aagataaaga acgttttgaa cgtgtgcgta atagcaccct
     4141 ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga taaggatcct aattggtaac
     4201 gaatcagaca attgacggct cgagggagta gcatagggtt tgcagaatcc ctgcttcgtc
     4261 catttgacag gcacattatg catcgatgat aagctgtcaa acatgagcag atcctctacg
     4321 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     4381 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc aaatatttta
     4441 tctgaggtgc ttcctcgctc
//
