LOCUS       Exported                4572 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 4572)
  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..4572
                     /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     2064..2142
                     /label=oGML0572_f
                     /note="create CYP029_cin_4-5, insert"
     RBS             2087..2115
                     /label=RBS
                     /note="strength = 39630"
     primer_bind     2092..2141
                     /label=oGML0589_f
                     /note="amplify CYP029"
     CDS             2116..3480
                     /codon_start=1
                     /product="CYP029"
                     /label=CYP029
                     /db_xref="UniProtKB/Swiss-Prot:B5GZC3"
                     /db_xref="Refseq:GI:197705857|GI:294329094|GI:490054986|EMB
                     L-CDS:EDY51669.1| 
                     EMBL-CDS:EFG10737.1|REFSEQ:WP_003957291.1"
                     /translation="MTTQKHDSSTTGNFTTGRAPGALPLIGHAGHLMRRPITFLESLST
                     YGDLVEIRIGTQRAYVPCHPELLRQVLTDDRTFDKGGVIIERFRSVLGNGLGTCPHSDH
                     RRQRRLMQPAFSKARLEEYSAVMRQEITALSGSWQEGQEIDAFAVLYGLGLRVVVRTLL
                     SSDLDDPTVEQVRHSFERTLHEFLPRTIIPQQIQQLPLAVNRRYHQELQALDEVVNRII
                     ADCRDAGLERGGLLATLTASHAGGGVPMDDSEIHDQVITMLLGGSESTGTTLTWALYLL
                     SQHPEIEMRLHEEVDTVLNGRPADWNDLPNLSLTHNIITETLRLYPPGWLLSRTTTKEV
                     ELAGTRMAPDTTIVFSPHVIQRQGDIHGCPDVFDPDRWSPDRTADLPRGNFVSFGGGPR
                     KCIGDTFGEVQSTLALSTIAARWKMECAPGSDIRPVAFDFAYRPRRLILRPRLR*"
     primer_bind     2571..2586
                     /label=seq1_CYP29
     primer_bind     3257..3273
                     /label=seq2_CYP29
     primer_bind     complement(3460..3480)
                     /label=oGML0572_r
                     /note="create CYP029_cin_4-5, insert"
     primer_bind     complement(3461..3478)
                     /label=oGML0589_r
                     /note="amplify CYP029"
     terminator      3524..3570
                     /label=L3S3P41
     misc_feature    3562
                     /label=deletion
     misc_feature    3566
                     /label=C to G
     misc_feature    3580..3583
                     /label=4bp_scar6
     promoter        3599..3680
                     /label=Constitutive Promoter + RBS (DNA2.0 Vector)
                     /note="Geneious type: promoter"
     promoter        3617..3622
                     /label=-35 promoter
                     /label=-35
     misc_feature    3618
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     promoter        3640..3645
                     /label=-10 promoter
                     /label=-10
     CDS             3681..4292
                     /label=CymR
     misc_feature    3978..3980
                     /label=E100D
                     /note="Geneious type: mutation"
     misc_feature    3980
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4008..4010
                     /label=S110
                     /note="Geneious type: mutation"
     misc_feature    4008
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4062..4064
                     /label=Q128L
                     /note="Geneious type: mutation"
     misc_feature    4063
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     misc_feature    4191..4193
                     /label=A171
                     /note="Geneious type: mutation"
     misc_feature    4191
                     /label=G
                     /note="Geneious type: Editing History Replacement"
     terminator      4302..4556
                     /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 tgtttaggcc ccccatataa
     2101 aattggagta tttttatgac tacacagaaa cacgattcgt ctacaaccgg gaatttcacc
     2161 actggccgcg ccccaggagc gctcccatta atcgggcacg caggccatct gatgcgccgt
     2221 cctatcacat ttctggaaag ccttagcacg tacggagatt tagtcgagat tcgtatcggt
     2281 acccaacgcg cgtatgtccc atgtcacccg gaactgttac gccaggttct cactgacgat
     2341 cggaccttcg ataaaggtgg agtaattatc gagcgttttc gttccgtgct gggaaacggc
     2401 ttgggaacat gtccacactc cgaccaccgt cggcagcgcc gtctgatgca gccggcattc
     2461 tcaaaagcgc gcctggaaga gtatagtgct gtgatgcggc aagaaatcac cgcgctctct
     2521 ggctcgtggc aagagggaca agagattgac gcctttgccg tgttgtacgg tctgggcctg
     2581 cgagtcgtag tgcgcacact gctgagctcc gatctggatg atccgaccgt tgaacaagtc
     2641 agacatagtt ttgagcgcac gttacacgag ttcctgccgc gcacaattat cccgcagcaa
     2701 atccagcaac tgccattagc cgtgaataga agataccacc aggaattgca agcgctggac
     2761 gaggtcgtta accgcatcat tgctgactgt cgtgacgctg gtttggaaag aggcggtctg
     2821 ctggccaccc tgacggcgag ccatgcagga ggcggagttc ctatggatga tagtgagatt
     2881 cacgatcagg tgatcaccat gctgttaggc ggatcggaaa gcaccggcac gaccctgact
     2941 tgggccctgt atctgctgag tcagcatccc gaaattgaaa tgcgtctgca tgaagaggtg
     3001 gataccgtcc tcaacggacg tccagcagat tggaacgatt tgccaaatct ctcacttacc
     3061 cacaatatca ttacggaaac gttgcgtctg tatccgccag gttggctgct gagccggacc
     3121 acaacaaaag aagttgaact ggccggaacc cggatggcac cggatacgac tattgtattt
     3181 tcgccccacg taatccaacg acagggtgat attcatggat gcccagatgt tttcgacccg
     3241 gaccgttggt cgcccgatag aacggcggac ctgccgcgtg gtaattttgt gagttttggc
     3301 ggcgggcccc gtaaatgtat tggtgacact tttggtgaag ttcagagcac cctggcactg
     3361 agtacgattg cagcgcgctg gaaaatggaa tgtgccccag ggagtgatat tcgtccggtt
     3421 gcctttgact ttgcatatcg tccgcgccgg cttatcctgc gccctcgcct ccgataataa
     3481 ggatgagctc tacaaataag cagaggtggt tgtgttgcga aaaaaaaaaa aaaaacaccc
     3541 taacgggtgt tttttttttt ttggtgtccc cacgtgtggc gctggagacc gtccaatggc
     3601 ggcgcgccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     3661 gtcaattcag ggtggtgaat atgagcccga aacgtcgtac ccaggcagaa cgtgcaatgg
     3721 aaacccaggg taaactgatt gcagcagcac tgggtgttct gcgtgaaaaa ggttatgcag
     3781 gttttcgtat tgcagatgtt ccgggtgcag ccggtgttag ccgtggtgca cagagccatc
     3841 attttccgac caaactggaa ctgctgctgg caacctttga atggctgtat gagcagatta
     3901 ccgaacgtag ccgtgcacgt ctggcaaaac tgaaaccgga agatgatgtt attcagcaga
     3961 tgctggatga tgcagcagat ttttttctgg atgatgattt tagcatcggc ctggatctga
     4021 ttgttgcagc agatcgtgat ccggcactgc gtgaaggtat tctgcgtacc gttgaacgta
     4081 atcgttttgt tgttgaagat atgtggctgg gtgtgctggt gagccgtggt ctgagccgtg
     4141 atgatgccga agatattctg tggctgattt ttaacagcgt tcgtggtctg acagttcgta
     4201 gcctgtggca gaaagataaa gaacgttttg aacgtgtgcg taatagcacc ctggaaattg
     4261 cacgtgaacg ttatgcaaaa ttcaaacgtt gataaggatc ctaattggta acgaatcaga
     4321 caattgacgg ctcgagggag tagcataggg tttgcagaat ccctgcttcg tccatttgac
     4381 aggcacatta tgcatcgatg ataagctgtc aaacatgagc agatcctcta cgccggacgc
     4441 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     4501 accgatgggg aagatcgggc tcgccacttc gggctcatga gcaaatattt tatctgaggt
     4561 gcttcctcgc tc
//
