LOCUS       sequence-236024-        2414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly CD part for ccdB (DNA gyrase toxin, 
            modified synonymously to remove BsaI cut site; ccdA/C79m is 
            antitoxin). Please see Supplemental Documents for annotated Genbank 
            file..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2414)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2414)
  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: C39m
FEATURES             Location/Qualifiers
     source          1..2414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..42
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(46..89)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    92..113
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             132..437
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     316..335
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    449..469
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(475..492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(646..1234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(726..745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1405..2196,2197..2265))
                     /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     2028..2047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2266..2370)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
       61 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagactagtg
      121 ggtctcaaat gatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
      181 ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg
      241 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
      301 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtg tccgttatcg
      361 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
      421 tgttctgggg aatataaagg tagagaccta ctagtagcgg ccgctgcagg cttcctcgct
      481 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg
      661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1081 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1201 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1321 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1381 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1441 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1501 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgcgacc cacgctcacc
     1561 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1621 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1681 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1741 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1801 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1861 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1921 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1981 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2041 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2101 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2161 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2221 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2281 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2341 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2401 ctaagaaacc atta
//
