LOCUS       Exported                4585 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 4585)
  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..4585
                     /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..2114
                     /label=RBS
                     /note="strength = 44869"
     CDS             2115..3494
                     /codon_start=1
                     /product="CYP021"
                     /label=CYP021
                     /db_xref="UniProtKB/Swiss-Prot:A0A1G8D6F5"
                     /db_xref="Refseq:EMBL-CDS:SDH53327.1"
                     /translation="MFNATGTTVTRPKVIPFYRAIPRLVKDPVTELARMGTEAGGRVVR
                     LDLGPFRPYLISHPDHVQQVLKTDWDNFVRAGMFWQPLQRLIGHSIMGEGESWASARKI
                     LQPLFTARYTASLAEDMADIIAERIVELDDFAQAGRPIDVEHEMTSIVNQAVIKVLFGG
                     RITRETAERLAPEFATCASSIGFRMLFPSAPYSIRVPGDRAFLRATKKIHEIVYPLIER
                     AQREKTQSKEVVSALLTVRLEQEGKADVRQVRDDLVNVYGAASENTAMSLLWLWSVLHD
                     HPEVNARLQEEIHRVVGDGPVRPSHVPEMHYLRMVLQELLRVYPSAWVLPRQAVADTEI
                     GGVRIKAGSQVLVSPYTTHRLEEFWPRPTEFDPERWAGDKTERRHRYSFIPFGAGPHVC
                     IGQHLFYVEASLLIANLFSRYRPALPYPQKLTPLRGGATLRPKEKLFLQLHPIERKRAA
                     *"
     primer_bind     2115..2134
                     /label=oGML0584_f
                     /note="amplify CYP021"
     primer_bind     2591..2606
                     /label=seq1_CYP21
     primer_bind     3299..3315
                     /label=seq2_CYP21
     primer_bind     complement(3476..3492)
                     /label=oGML0584_r
                     /note="amplify CYP021"
     terminator      3537..3583
                     /label=L3S3P41
     misc_feature    3575
                     /label=deletion
     misc_feature    3579
                     /label=C to G
     misc_feature    3593..3596
                     /label=4bp_scar6
     promoter        3612..3693
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3630..3635
                     /label=-35 promoter
                     /label=-35
     misc_feature    3631
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3653..3658
                     /label=-10 promoter
                     /label=-10
     CDS             3694..4305
                     /label=CymR
     misc_feature    3991..3993
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    3993
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4021..4023
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    4021
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4075..4077
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    4076
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4204..4206
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4204
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4315..4569
                     /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 tgtttaggat atagtagcac
     2101 gaagggggta cttaatgttt aacgccactg gcacgaccgt aacacgcccc aaagttatcc
     2161 cattctacag agccattcca cgtttggtta aagatccagt gaccgagctt gctcgtatgg
     2221 gaacggaagc agggggacgt gtggtgcggt tagaccttgg tccgtttcgt ccgtatctga
     2281 tcagccaccc ggaccatgtg caacaggtac tgaaaacgga ttgggataat tttgtccgtg
     2341 cgggcatgtt ctggcaacct ctgcagcgcc tcatcgggca ctctataatg ggagaaggcg
     2401 agtcgtgggc aagtgcgcgc aaaatcctcc agcccctgtt tacggcacgc tacaccgcgt
     2461 ccctggctga agatatggcg gatatcatag ccgagcgcat tgtggaactg gatgattttg
     2521 cgcaggcagg ccgcccgatc gatgttgaac atgaaatgac gtctattgtt aatcaggcgg
     2581 ttattaaagt tctgttcggt ggtcgcataa ctcgagaaac agcagaacgc ttagcacccg
     2641 agttcgccac atgcgccagt tctatagggt tccgtatgct gttcccgtcg gctccttact
     2701 cgatccgcgt accaggagac agagccttcc tgcgtgccac gaagaaaatc catgagatcg
     2761 tttatcccct gatcgagcgt gctcagcgtg aaaagacgca gagcaaggaa gtggtgtcag
     2821 ccttactgac ggtgcgcctg gaacaggaag gtaaggccga cgttcgccaa gttcgtgatg
     2881 acttagtgaa tgtatatggt gcggcaagcg agaataccgc gatgtccctg ctgtggctgt
     2941 ggagtgtcct gcacgaccat ccggaagtaa acgcaagatt gcaggaggag attcaccgag
     3001 ttgtgggtga tggtccggtc cgtccttccc acgtcccgga aatgcattac ctgcgcatgg
     3061 tgcttcaaga actgctgcgc gtgtatcctt ctgcttgggt tttacctcgc caagcggtag
     3121 cggatacgga aattggtggt gtccgcatta aagccggaag ccaggtactg gttagcccct
     3181 acactacaca tagactcgaa gaattttggc ctcgcccgac cgaatttgat ccagagagat
     3241 gggccgggga taagacggaa agacgccatc gttactcctt tatcccgttt ggcgctggtc
     3301 cgcatgtctg cattggccag catttattct acgtggaggc ctcgctgttg attgccaatc
     3361 ttttctctcg ctaccgcccg gcgctgccgt acccgcagaa acttacgcca cttcgtggcg
     3421 gggcgacgct ccgcccgaaa gaaaagctgt ttttgcaact gcatccgatc gaacgcaagc
     3481 gcgccgcgta ataaggatga gctctacaaa taagcagagg tggttgtgtt gcgaaaaaaa
     3541 aaaaaaaaca ccctaacggg tgtttttttt tttttggtgt ccccacgtgt ggcgctggag
     3601 accgtccaat ggcggcgcgc catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag
     3661 cgcccggaag agagtcaatt cagggtggtg aatatgagcc cgaaacgtcg tacccaggca
     3721 gaacgtgcaa tggaaaccca gggtaaactg attgcagcag cactgggtgt tctgcgtgaa
     3781 aaaggttatg caggttttcg tattgcagat gttccgggtg cagccggtgt tagccgtggt
     3841 gcacagagcc atcattttcc gaccaaactg gaactgctgc tggcaacctt tgaatggctg
     3901 tatgagcaga ttaccgaacg tagccgtgca cgtctggcaa aactgaaacc ggaagatgat
     3961 gttattcagc agatgctgga tgatgcagca gatttttttc tggatgatga ttttagcatc
     4021 ggcctggatc tgattgttgc agcagatcgt gatccggcac tgcgtgaagg tattctgcgt
     4081 accgttgaac gtaatcgttt tgttgttgaa gatatgtggc tgggtgtgct ggtgagccgt
     4141 ggtctgagcc gtgatgatgc cgaagatatt ctgtggctga tttttaacag cgttcgtggt
     4201 ctgacagttc gtagcctgtg gcagaaagat aaagaacgtt ttgaacgtgt gcgtaatagc
     4261 accctggaaa ttgcacgtga acgttatgca aaattcaaac gttgataagg atcctaattg
     4321 gtaacgaatc agacaattga cggctcgagg gagtagcata gggtttgcag aatccctgct
     4381 tcgtccattt gacaggcaca ttatgcatcg atgataagct gtcaaacatg agcagatcct
     4441 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     4501 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcaaata
     4561 ttttatctga ggtgcttcct cgctc
//
