LOCUS       sequence-236034-        2822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly CD part for cI857 (temperature-sensitive 
            lambda repressor that represses at 30C but not at 37C; see 
            https://doi.org/10.1016/S0022-2836(66)80269-3. Use with P9m)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2822)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2822)
  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"
COMMENT     Sequence Label: C64m
FEATURES             Location/Qualifiers
     source          1..2822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..46
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(50..93)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    96..117
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             136..849
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     misc_feature    861..881
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(887..904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1058..1646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1138..1157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1817..2608,2609..2677))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2440..2459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2678..2782)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa
       61 aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct tctagagact
      121 agtgggtctc aaatgatgag cacaaaaaag aaaccattaa cacaagagca gcttgaggac
      181 gcacgtcgcc ttaaagcaat ttatgaaaaa aagaaaaatg aacttggctt atcccaggaa
      241 tctgtcgcag acaagatggg gatggggcag tcaggcgttg gtgctttatt taatggcatc
      301 aatgcattaa atgcttataa cgccgcattg cttacaaaaa ttctcaaagt tagcgttgaa
      361 gaatttagcc cttcaatcgc cagagaaatc tacgagatgt atgaagcggt tagtatgcag
      421 ccgtcactta gaagtgagta tgagtaccct gttttttctc atgttcaggc agggatgttc
      481 tcacctaagc ttagaacctt taccaaaggt gatgcggaga gatgggtaag cacaaccaaa
      541 aaagccagtg attctgcatt ctggcttgag gttgaaggta attccatgac cgcaccaaca
      601 ggctccaagc caagctttcc tgacggaatg ttaattctcg ttgaccctga gcaggctgtt
      661 gagccaggtg atttctgcat agccagactt gggggtgatg agtttacctt caagaaactg
      721 atcagggata gcggtcaggt gtttttacaa ccactaaacc cacagtaccc aatgatccca
      781 tgcaatgaga gttgttccgt tgtggggaaa gttatcgcta gtcagtggcc tgaagagacg
      841 tttggctgaa ggtagagacc tactagtagc ggccgctgca ggcttcctcg ctcactgact
      901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc cgcccccctg
     1081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1861 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1921 gcttaccatc tggccccagt gctgcaatga taccgcgcga cccacgctca ccggctccag
     1981 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2041 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2101 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2161 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2221 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2281 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2341 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2401 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2461 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2521 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2581 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2641 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2701 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2761 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2821 cc
//
