LOCUS       sequence-231078-        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"
     CDS             5..310
                     /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     189..208
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    322..342
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(348..365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(519..1107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(599..618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1278..2069,2070..2138))
                     /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     1901..1920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2139..2243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2311..2329
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2333..2376)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2379..2400
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aatgatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
       61 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
      121 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
      181 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtgtccgtta tcggggaaga
      241 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
      301 gggaatataa aggtagagac ctactagtag cggccgctgc aggcttcctc gctcactgac
      361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct
      541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1381 ggcttaccat ctggccccag tgctgcaatg ataccgcgcg acccacgctc accggctcca
     1441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2281 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     2341 taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag
     2401 actagtgggt ctca
//
