LOCUS       Exported                4655 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 4655)
  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..4655
                     /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..2109
                     /label=RBS
                     /note="strength = 50703"
     CDS             2110..3564
                     /codon_start=1
                     /product="CYP022"
                     /label=CYP022
                     /db_xref="UniProtKB/Swiss-Prot:A0A1Z4J4D1"
                     /db_xref="Refseq:EMBL-CDS:BAY49125.1"
                     /translation="MEIKEGYFKQQQTLKNADTAQEHQINRPSPFDPSPTPKTCPVVRG
                     YPFVGVLPELLQNPLRLFQRAVKDHPGEIVALKLGPIQAHLVTHPDHVKYVLNDNWQNF
                     GKGEVMWKPARQVVGNGLITSHGESWFRNRRLMQPMFAANQANALTELSVKLIAEQLAR
                     FETTANSEPIDIKNEMALLTQTVILGVIFSSSIDRSKAETLTEALKVLVRETSRRMFLC
                     FLPKQFPIPGERAFRNAIQTTDDTILSIIRARRWSSLNYGDLFERLLFSRDEESKTGMN
                     DQQLRDESITLFLAGYETTSTALTWVWYLLDKHPEVQSKLRAEIDAVIGKRQPTCADVA
                     KLQYTKMVIQEVMRLYPPVLFLARVVEADDVIDGYPLKAGTTVVLCKYLTHRLPEFWDQ
                     PEVFDPERFASEYSTQRHSYSYFPFGGGPRYCIGKHLALLEMQLIVAMMVQKYRLRCVP
                     GHPVEASPKIILHPRYGLPMTVEPV*"
     primer_bind     2110..2137
                     /label=oGML0585_f
                     /note="amplify CYP022"
     primer_bind     2583..2597
                     /label=seq1_CYP22
     primer_bind     3272..3288
                     /label=seq2_CYP22
     primer_bind     complement(3539..3562)
                     /label=oGML0585_r
                     /note="amplify CYP022"
     terminator      3607..3653
                     /label=L3S3P41
     misc_feature    3649
                     /label=C to G
     misc_feature    3663..3666
                     /label=4bp_scar6
     promoter        3682..3763
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3700..3705
                     /label=-35 promoter
                     /label=-35
     misc_feature    3701
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3723..3728
                     /label=-10 promoter
                     /label=-10
     CDS             3764..4375
                     /label=CymR
     misc_feature    4061..4063
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    4063
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4091..4093
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    4091
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4145..4147
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    4146
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4274..4276
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4274
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4385..4639
                     /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 tgtttaggac ctagcagaag
     2101 gagaataaaa tggaaatcaa ggaaggctat tttaagcagc aacaaaccct caaaaatgcc
     2161 gataccgccc aggaacacca aattaaccgc ccatcgccgt tcgatccgtc ccccaccccg
     2221 aaaacctgtc cagtcgtgcg tggttaccca tttgtaggcg tactgccaga actgctgcag
     2281 aacccgctgc gcctgtttca gcgggcagtg aaagaccatc cgggagaaat tgttgccctg
     2341 aaactggggc caattcaagc ccatttagta acacacccgg atcacgtgaa atatgtgttg
     2401 aatgataatt ggcagaattt tgggaaaggt gaagtcatgt ggaaaccagc ccgtcaggtc
     2461 gtcggtaacg gcctgatcac aagccacggc gaaagttggt ttcgcaatcg cagactgatg
     2521 cagccgatgt ttgcggcgaa tcaggctaac gcgctgaccg agctgtcagt taaacttatc
     2581 gctgaacagc tcgcacgttt tgaaaccaca gcaaattcag aacctatcga cattaagaac
     2641 gaaatggcac tgttaacgca gacggtgatc ctgggcgtta tattctcctc tagcattgat
     2701 cgttccaaag cagaaacgct caccgaagct ctgaaagttc tggttcgaga aaccagtcgt
     2761 agaatgtttc tgtgttttct gccaaaacag tttccgatcc cgggtgaacg cgcgttccgc
     2821 aatgcgattc aaactacaga tgatacaatt ctgtcaatca ttcgcgcccg gcgttggtct
     2881 agccttaact acggagactt gtttgagaga ttactgttta gccgtgatga ggaatcaaaa
     2941 accggaatga acgaccagca actgcgggac gagagcatta cactgttcct cgcaggatat
     3001 gaaaccacga gcaccgcact gacgtgggtt tggtatttgt tggacaaaca ccccgaagta
     3061 cagagtaaac tccgcgccga aattgacgct gttattggga aacgccagcc cacttgtgct
     3121 gatgtagcga aactgcagta caccaaaatg gttattcaag aagtgatgcg actctacccg
     3181 cccgtgctgt ttctggcgcg ggtagtggaa gctgacgacg tgattgatgg ctacccgctg
     3241 aaagctggca caaccgtcgt cctgtgtaag tacctgactc atcgtctgcc tgaattttgg
     3301 gatcaaccgg aagtctttga tccagagcgt ttcgcatcag aatatagtac acagcgtcat
     3361 tcgtattcct attttccttt cggcggtggt ccacgttatt gcattgggaa acatttggca
     3421 cttctcgaga tgcagcttat tgtcgctatg atggtacaaa agtatcgttt gcgttgcgtc
     3481 cccggccatc cggtggaggc gagtcctaaa attatcctgc accctcgcta cggcctcccg
     3541 atgactgtgg aaccggttta ataaggatga gctctacaaa taagcagagg tggttgtgtt
     3601 gcgaaaaaaa aaaaaaaaca ccctaacggg tgtttttttt tttttggtgt ccccacgtgt
     3661 ggcgctggag accgtccaat ggcggcgcgc catcgaatgg tgcaaaacct ttcgcggtat
     3721 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatatgagcc cgaaacgtcg
     3781 tacccaggca gaacgtgcaa tggaaaccca gggtaaactg attgcagcag cactgggtgt
     3841 tctgcgtgaa aaaggttatg caggttttcg tattgcagat gttccgggtg cagccggtgt
     3901 tagccgtggt gcacagagcc atcattttcc gaccaaactg gaactgctgc tggcaacctt
     3961 tgaatggctg tatgagcaga ttaccgaacg tagccgtgca cgtctggcaa aactgaaacc
     4021 ggaagatgat gttattcagc agatgctgga tgatgcagca gatttttttc tggatgatga
     4081 ttttagcatc ggcctggatc tgattgttgc agcagatcgt gatccggcac tgcgtgaagg
     4141 tattctgcgt accgttgaac gtaatcgttt tgttgttgaa gatatgtggc tgggtgtgct
     4201 ggtgagccgt ggtctgagcc gtgatgatgc cgaagatatt ctgtggctga tttttaacag
     4261 cgttcgtggt ctgacagttc gtagcctgtg gcagaaagat aaagaacgtt ttgaacgtgt
     4321 gcgtaatagc accctggaaa ttgcacgtga acgttatgca aaattcaaac gttgataagg
     4381 atcctaattg gtaacgaatc agacaattga cggctcgagg gagtagcata gggtttgcag
     4441 aatccctgct tcgtccattt gacaggcaca ttatgcatcg atgataagct gtcaaacatg
     4501 agcagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     4561 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     4621 tgagcaaata ttttatctga ggtgcttcct cgctc
//
