LOCUS       sequence-231087-        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"
     CDS             5..718
                     /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    730..750
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(756..773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(927..1515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1007..1026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1686..2477,2478..2546))
                     /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     2309..2328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2547..2651)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2719..2737
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2741..2784)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2787..2808
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aatgatgagc acaaaaaaga aaccattaac acaagagcag cttgaggacg cacgtcgcct
       61 taaagcaatt tatgaaaaaa agaaaaatga acttggctta tcccaggaat ctgtcgcaga
      121 caagatgggg atggggcagt caggcgttgg tgctttattt aatggcatca atgcattaaa
      181 tgcttataac gccgcattgc ttacaaaaat tctcaaagtt agcgttgaag aatttagccc
      241 ttcaatcgcc agagaaatct acgagatgta tgaagcggtt agtatgcagc cgtcacttag
      301 aagtgagtat gagtaccctg ttttttctca tgttcaggca gggatgttct cacctaagct
      361 tagaaccttt accaaaggtg atgcggagag atgggtaagc acaaccaaaa aagccagtga
      421 ttctgcattc tggcttgagg ttgaaggtaa ttccatgacc gcaccaacag gctccaagcc
      481 aagctttcct gacggaatgt taattctcgt tgaccctgag caggctgttg agccaggtga
      541 tttctgcata gccagacttg ggggtgatga gtttaccttc aagaaactga tcagggatag
      601 cggtcaggtg tttttacaac cactaaaccc acagtaccca atgatcccat gcaatgagag
      661 ttgttccgtt gtggggaaag ttatcgctag tcagtggcct gaagagacgt ttggctgaag
      721 gtagagacct actagtagcg gccgctgcag gcttcctcgc tcactgactc gctgcgctcg
      781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      901 cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac
      961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1081 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1201 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1261 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1321 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1381 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1441 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1501 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1561 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1621 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1681 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1741 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1801 ggccccagtg ctgcaatgat accgcgcgac ccacgctcac cggctccaga tttatcagca
     1861 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1921 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1981 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2041 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2101 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2161 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2221 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2281 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2341 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2401 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2461 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2521 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2581 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2641 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2701 atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc
     2761 ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctagagac tagtgggtct
     2821 ca
//
