LOCUS       sequence-12387-e        8090 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 8090)
  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 8090)
  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: pBCN28-R4R3
FEATURES             Location/Qualifiers
     source          1..8090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2994..3013)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3592..3612
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4274..4296
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4334..4352)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4420..4524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4525..4593,4594..5385)
                     /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(4743..4762)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5556..6144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6045..6064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6270..6292
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6289..6305
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6289..6305
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6325..6448
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6489..6794)
                     /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(6591..6610)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(7114..7794)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     7709..7728
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7848..7854,7855..7872,7873..7878))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(7903..8026)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(8034..8051)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8034..8050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8043..8065)
                     /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 gccccatttt atatctgagt
      181 agtatccttt gctttaaatg tccataaaac taattttata atcaataaaa caacgtttgt
      241 aaatcaactg agtttacaag tagagacatt gagggatact ttcactatgc taaagtgata
      301 atcgaccaaa taataacttc actttggtat ttattcctgt cttataaatg ttatgtatga
      361 attaaattca tatgcatatg gctcactctg acaattttta ataatcttcc agatcaatat
      421 tgactaccga tgcgggtggt cttttgcttt gaattctgct gaactttaca ccccgaacag
      481 caatgtgtgc ttcagcctaa aaaaaagtaa gtgtgttaat cagtgccccc gattcttcat
      541 tttttgcccc tctctcccgt ttcgtcggca aaagaagaga aaataaagat aagtctcaag
      601 ataggttggt aatcgctaaa gtggttgtgt ggataagagt agcaaaatgg caggaagagc
      661 actttgcgcg cacacactgt actcattgtt ctggataaaa ttctctcgtt gtttgccgtc
      721 ggatgtctgc ctctctgcat tgagccggct tcttcactat ctttagttaa cctaaaatgc
      781 cgtttctttt ctcgtatccc cactatcccg ttgaggttct ctgctctctt cgctccctac
      841 cgccagcgag caactatccg tgggggcgcc ttgctcggaa gatggggggg aagaaagaag
      901 atttttgcta tttgcacttg agaaagagac ttttcctgcg tcgatggtta gagaacagtg
      961 tgcagacact tttcagctac ctagaattac aattggatat ccccgcctcc caatccaccc
     1021 acccagggaa aaagaagggc tcgccgaaaa tcaaagttat ctccaggctc gcgcatccca
     1081 ccgagcggtt gacttctctc caccactttt cattttaacc ctcgatcgtc agacacagaa
     1141 atggtctcaa agggtgaaga agataacatg gcaattatta aagagtttat gcgtttcaag
     1201 gtgcatatgg agggatctgt caatgggcat gagtttgaaa ttgaaggtga aggagaaggc
     1261 cgaccatatg agggaacaca aaccgcaaaa ctaaaggtaa gtttaaacat atatatacta
     1321 actaaccctg attatttaaa ttttcaggta actaaaggcg gaccattacc attcgcctgg
     1381 gacatcctct ctccacagtt catgtatgga agtaaagctt atgttaaaca tccggcagat
     1441 ataccagatt atttgaaact ttcattcccg gagggtttta agtgggaacg cgtaatgaat
     1501 tttgaagacg gaggagttgt tacagtgacg caagactcaa ggtaagttta aacagttcgg
     1561 tactaactaa ccatacatat ttaaattttc agcctccaag atggagaatt tatttataaa
     1621 gtcaaacttc gaggaacgaa tttcccctcg gatggacctg ttatgcagaa gaagactatg
     1681 ggatgggaag cttcaagtga aagaatgtac cctgaagacg gtgctcttaa gggagagatt
     1741 aaacaacgtc ttaaattgaa agatggagga cattacgatg ctgaggtaag tttaaacatg
     1801 attttactaa ctaactaatc tgatttaaat tttcaggtga agacaactta caaagccaaa
     1861 aaaccagttc agctgccagg agcgtacaat gttaatatta aactggatat cacctcccac
     1921 aacgaggatt acactatcgt tgagcaatat gaaagagctg aagggcggca ctcgacaggt
     1981 ggcatggatg aattgtataa gtaggctccg catcggccgc tgtcatcaga tcgccatctc
     2041 gcgcccgtgc ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct
     2101 ccctgtgctc ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt
     2161 ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta
     2221 attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc
     2281 ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt
     2341 ttgaaacatc atagaaaaaa ccgcacacag ggccccccct cgaggggaga aaatgcggag
     2401 agaagaaccc agcatcttaa taaaattata taattttgaa taaaatcaaa ttaaaaataa
     2461 gattaattga aacacaacaa tagaatgcag aaaattactc ataagaaata atttattttg
     2521 aaagaagttg tgtcgattta cgataaaaac tacacgcaaa cgaggctccg cgtttacacc
     2581 gtttggcaat ggagcgcgct tgctattaga ttttaggcca gctcaacgaa agtttaattt
     2641 atttttcttc tttttttcca tttttttgag taaaatatcg aattttttgt attaattctg
     2701 taaattttat aatttgcaac tattttggcg tattctgctt gtttttatca gcgaattttt
     2761 atgcatttca gtcaaaagtt tgagcaggaa aaatgacatt taggactttg taaaattgta
     2821 ttcttaaatt tgcttgaaaa cctacgtgta caattgattt tattgaattt ttcttacgtt
     2881 tcccgttgta tcttattgta tttaactttc atcataaaat aatttatttc caggtaaaat
     2941 atccgacgct ctcgtggatc cccgggattg gctcagaaaa aaaggtaccg accatgaccg
     3001 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     3061 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     3121 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     3181 aggtgtgggt cggtaagttt aaacatatat atactaacta accctgatta tttaaatttt
     3241 cagcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     3301 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     3361 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     3421 ccaccgtcgg cgtctcgccc gacgtaagtt taaacatgat tttactaact aactaatctg
     3481 atttaaattt tcagcaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     3541 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     3601 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     3661 gcacctggtg catgacccgc aagcccggtg cctgatgagc tagccggcca tacaagtaat
     3721 ccggatgatc gacgccaacg tcgttgaatt ttcaaatttt aaatactgaa tatttgtttt
     3781 ttttcctatt atttatttat tctctttgtg ttttttttct tgctttctaa aaaattaatt
     3841 caatccaaat ctaaacattt ttttttctct ttccgtctcc caattcgtat tccgctcctc
     3901 tcatctgaac acaatgtgca agtttattta tcttctcgct ttcatttcat taggacgtgg
     3961 ggggaattgg tggaaggggg aaacacacaa aaggatgatg gaaatgaaat aaggacacac
     4021 aatatgcaac aacattcaat tcagaaatat ggaggaaggt ttaaaagaaa acataaaaat
     4081 atatagagga ggaaggaaaa ctagttctag agcggccgcc accgcggtgg agctcccata
     4141 tggtgcactc tcagtgcaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4201 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4261 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4321 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4381 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4441 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4501 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4561 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4621 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4681 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4741 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4801 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4861 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4921 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4981 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5041 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5101 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5161 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5221 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5281 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5341 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5401 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5461 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5521 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5581 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5641 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5701 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5761 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5821 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5881 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5941 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6001 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6061 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6121 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggtc
     6181 ttttgctggc ctttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6241 gtattaccgc ctttgagtga gctgatacca gcggataaca atttcacaca ggaaacagct
     6301 atgaccatga ttacgccaag ctatcaactt tgtatagaaa agttgaacga gaaacgtaaa
     6361 atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact
     6421 gtaaaacaca acatatccag tcactatggt cgacctgcag actggctgtg tataagggag
     6481 cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg
     6541 tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg
     6601 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggggact
     6661 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca
     6721 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     6781 accttaaact gcatttcacc agcccctgtt ctcgtcggca aaagagccgt tcatttcaat
     6841 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag
     6901 acgacgggct tcattctgca tggttgtgct taccgaaccg gagatattga catcatatat
     6961 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca
     7021 tacctctttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat
     7081 cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcct ctagattacg
     7141 ccccgcctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     7201 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     7261 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     7321 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     7381 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     7441 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     7501 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     7561 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     7621 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     7681 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     7741 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     7801 ttccttagct cctgaaaatc tcgacggatc ctaactcaaa atccacacat tatacgagcc
     7861 ggaagcataa agtgtaaagc ctggggtgcc taatgcggcc gccatagtga ctggatatgt
     7921 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     7981 tttatatcat tttacgtttc tcgttcaact ttattataca tagttgataa ttcactggcc
     8041 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
//
