LOCUS       sequence-209531-        3045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MIDAS Level-1 source vector for cloning of modules (synthetic 
            biology parts).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3045)
  AUTHORS   van Dolleweerd CJ, Kessans SA, Van de Bittner KC, Bustamante LY, 
            Bundela R, Scott B, Nicholson MJ, Parker EJ
  TITLE     MIDAS: A Modular DNA Assembly System for Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2018 Apr 20;7(4):1018-1029. doi: 
            10.1021/acssynbio.7b00363. Epub 2018 Apr 12.
  PUBMED    29620866
REFERENCE   2  (bases 1 to 3045)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00363"; journalName: "ACS Synth Biol"; date: 
            "2018-04-20- 20"; volume: "7"; issue: "4"; pages: "1018-1029"
COMMENT     Sequence Label: pML1
FEATURES             Location/Qualifiers
     source          1..3045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(20..193)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     142..164
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     156..173
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     157..173
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(189..205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(189..205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(202..224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    213..229
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(237..243,244..261,262..267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    282..303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(441..458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(612..1200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(692..711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1672..2463
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(2555..3010)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2692..2713)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2904..2923
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 attaccaggc aaagcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
       61 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
      121 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaatccg
      181 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      301 ttaattgcgt tgcgccgtct ctagacctcg agggcatcac tgcccgcttt ccagtcggga
      361 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctaggtcgt tcggctgcgg
      481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      781 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1021 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaactaaaa ctcacgttaa
     1261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag accggacgtc
     1381 gatatccgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     1441 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     1501 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     1561 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1621 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa
     1681 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1741 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1801 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1861 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1921 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1981 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     2041 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     2101 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2161 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2221 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2281 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgta
     2341 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2401 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2461 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2521 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taatcgttaa tattttgtta
     2581 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2641 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2701 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2761 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2821 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2881 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2941 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3001 cagggcgcgt cccattcgcc agtgtccatg gtctcgtgag acgtt
//
