LOCUS       sequence-12385-e        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  AUTHORS   Semple JI, Garcia-Verdugo R, Lehner B
  TITLE     Rapid selection of transgenic C. elegans using antibiotic 
            resistance.
  JOURNAL   Nat Methods. 2010 Sep;7(9):725-7. doi: 10.1038/nmeth.1495. Epub 2010
            Aug 22.
  PUBMED    20729840
REFERENCE   2  (bases 1 to 5879)
  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"; doi: 
            "10.1038/nmeth.1495"; journalName: "Nat Methods"; date: "2010-09"; 
            volume: "7"; issue: "9"; pages: "725-7"
COMMENT     Sequence Label: pBCN21-R4R3
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(783..802)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1381..1401
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2063..2085
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2123..2141)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2209..2313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2314..2382,2383..3174)
                     /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     complement(2532..2551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3345..3933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3834..3853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4059..4081
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4078..4094
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4078..4094
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4114..4237
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4278..4583)
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(4380..4399)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4903..5583)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5498..5517
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5637..5643,5644..5661,5662..5667))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5692..5815)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5823..5840)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5823..5839)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5832..5854)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
       61 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
      121 ccttacgcat ctgtgcggta tttcacaccg catatgccgg gccccccctc gaggggagaa
      181 aatgcggaga gaagaaccca gcatcttaat aaaattatat aattttgaat aaaatcaaat
      241 taaaaataag attaattgaa acacaacaat agaatgcaga aaattactca taagaaataa
      301 tttattttga aagaagttgt gtcgatttac gataaaaact acacgcaaac gaggctccgc
      361 gtttacaccg tttggcaatg gagcgcgctt gctattagat tttaggccag ctcaacgaaa
      421 gtttaattta tttttcttct ttttttccat ttttttgagt aaaatatcga attttttgta
      481 ttaattctgt aaattttata atttgcaact attttggcgt attctgcttg tttttatcag
      541 cgaattttta tgcatttcag tcaaaagttt gagcaggaaa aatgacattt aggactttgt
      601 aaaattgtat tcttaaattt gcttgaaaac ctacgtgtac aattgatttt attgaatttt
      661 tcttacgttt cccgttgtat cttattgtat ttaactttca tcataaaata atttatttcc
      721 aggtaaaata tccgacgctc tcgtggatcc ccgggattgg ctcagaaaaa aaggtaccga
      781 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
      841 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
      901 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
      961 acatcggcaa ggtgtgggtc ggtaagttta aacatatata tactaactaa ccctgattat
     1021 ttaaattttc agcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     1081 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     1141 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     1201 ggttcctggc caccgtcggc gtctcgcccg acgtaagttt aaacatgatt ttactaacta
     1261 actaatctga tttaaatttt cagcaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     1321 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     1381 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     1441 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatgagct agccggccat
     1501 acaagtaatc cggatgatcg acgccaacgt cgttgaattt tcaaatttta aatactgaat
     1561 atttgttttt tttcctatta tttatttatt ctctttgtgt tttttttctt gctttctaaa
     1621 aaattaattc aatccaaatc taaacatttt tttttctctt tccgtctccc aattcgtatt
     1681 ccgctcctct catctgaaca caatgtgcaa gtttatttat cttctcgctt tcatttcatt
     1741 aggacgtggg gggaattggt ggaaggggga aacacacaaa aggatgatgg aaatgaaata
     1801 aggacacaca atatgcaaca acattcaatt cagaaatatg gaggaaggtt taaaagaaaa
     1861 cataaaaata tatagaggag gaaggaaaac tagttctaga gcggccgcca ccgcggtgga
     1921 gctcccatat ggtgcactct cagtgcaatc tgctctgatg ccgcatagtt aagccagccc
     1981 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     2041 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     2101 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2161 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2221 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2281 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2341 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2401 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2461 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2521 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2581 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2641 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2701 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2761 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2821 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2881 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2941 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3001 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3061 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3121 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3181 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3241 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3301 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3361 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3421 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3481 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3541 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3601 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3661 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3721 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3781 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3841 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3901 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3961 ttcctggtct tttgctggcc tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4021 tggataaccg tattaccgcc tttgagtgag ctgataccag cggataacaa tttcacacag
     4081 gaaacagcta tgaccatgat tacgccaagc tatcaacttt gtatagaaaa gttgaacgag
     4141 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     4201 cataatactg taaaacacaa catatccagt cactatggtc gacctgcaga ctggctgtgt
     4261 ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca
     4321 ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc
     4381 atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca
     4441 cgggggactt tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg
     4501 ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta
     4561 taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcggcaa aagagccgtt
     4621 catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc
     4681 ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accgaaccgg agatattgac
     4741 atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc
     4801 ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat attcttatac
     4861 cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatcctc
     4921 tagattacgc cccgcctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     4981 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     5041 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     5101 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     5161 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     5221 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     5281 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     5341 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     5401 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     5461 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     5521 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     5581 cattttagct tccttagctc ctgaaaatct cgacggatcc taactcaaaa tccacacatt
     5641 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgcggccg ccatagtgac
     5701 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
     5761 atattgatat ttatatcatt ttacgtttct cgttcaactt tattatacat agttgataat
     5821 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaa
//
