LOCUS       sequence-91667-e        4503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses optimized blasticidin S resistance gene (BSD) for C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4503)
  AUTHORS   Kim H, Ishidate T, Ghanta KS, Seth M, Conte D Jr, Shirayama M, Mello
            CC
  TITLE     A Co-CRISPR Strategy for Efficient Genome Editing in Caenorhabditis 
            elegans.
  JOURNAL   Genetics. 2014 May 30. pii: genetics.114.166389.
  PUBMED    24879462
REFERENCE   2  (bases 1 to 4503)
  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"; journalName: "Genetics. 
            2014 May 30. pii: genetics.114.166389."
COMMENT     Sequence Label: B1979
FEATURES             Location/Qualifiers
     source          1..4503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2315..2335)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2315..2333)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2354..2370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2354..2370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2367..2389)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2378..2394
                     /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(2402..2408,2409..2426,2427..2432))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2447..2468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2585..2602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2756..3344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2836..2855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3515..4306,4307..4375))
                     /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     4138..4157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4376..4480)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaggaaactg ctcgagggga gaaaatgcgg agagaagaac
      721 ccagcatctt aataaaatta tataattttg aataaaatca aattaaaaat aagattaatt
      781 gaaacacaac aatagaatgc agaaaattac tcataagaaa taatttattt tgaaagaagt
      841 tgtgtcgatt tacgataaaa actacacgca aacgaggctc cgcgtttaca ccgtttggca
      901 atggagcgcg cttgctatta gattttaggc cagctcaacg aaagtttaat ttatttttct
      961 tctttttttc catttttttg agtaaaatat cgaatttttt gtattaattc tgtaaatttt
     1021 ataatttgca actattttgg cgtattctgc ttgtttttat cagcgaattt ttatgcattt
     1081 cagtcaaaag tttgagcagg aaaaatgaca tttaggactt tgtaaaattg tattcttaaa
     1141 tttgcttgaa aacctacgtg tacaattgat tttattgaat ttttcttacg tttcccgttg
     1201 tatcttattg tatttaactt tcatcataaa ataatttatt tccaggtaaa atgggatccg
     1261 ctaaaccact ttcacaagag gagtcaacac tcatcgagcg tgctaccgca actatcaaca
     1321 gtatcccaat ttcggaagat tattcggttg ctagtgctgc actctcatct gatggacgta
     1381 ttttcacagg agtcaacgtt taccatttta ctggaggacc atgtgccgaa ctcgttgtgc
     1441 tgggaaccgc cgctgcagcc gctgcaggta agagattttt taaaaattta ttttttacac
     1501 tgttttttct caggaaactt gacttgcatt gtggcaatcg gaaatgagaa cagaggaatc
     1561 ctttccccat gtggaagatg ccgacaagtg cttttggatt tgcacccagg aattaaagca
     1621 atcgtcaagg attctgatgg acaaccaact gctgtgggaa tcagagaact gctgccatcg
     1681 ggatacgtgt gggagggagg atcctaaatg acgtttgtta cgttgttgtt atgaatataa
     1741 agtgttacag cttatcaaca cgttaaaatt ttttattcaa aaaaaaagtg aaaaatcaag
     1801 acaattagga gttgttcttt gtgaagcact caatccatct tttctggaaa aaataaatat
     1861 gtaaaattta gattagaaat cctaccttgt cgtcgtcaga tgcatgttca tctaaatata
     1921 aatcaatatc tgccttcaaa gaatcgattt cggttttttt ggcacgcttg ttcgagttag
     1981 caattttaat cattgcctga aaaatttttc aatatctaag aagtcgtttt aaaaataccg
     2041 aatgttcgtc gattacactt tgaacgccat caatggtttc ttttgaaatt ccacattttg
     2101 tcaacatttg aatcgggaat ccaccacctc cagagcaaat tgatacgaga agaaaggaca
     2161 gaatcaaaaa tttcatgttg aaatagcact caaatcggct caacgttgaa tgggcatgca
     2221 ttcgacgaaa ggtcgatcac gacattttag gttcagttga aaccggttat agggcgtcga
     2281 cctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     2341 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2401 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3181 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3781 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4381 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4441 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4501 cac
//
