LOCUS       sequence-12389-e        5549 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 5549)
  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 5549)
  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: pBCN25-R4R3
FEATURES             Location/Qualifiers
     source          1..5549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             783..1157
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     1733..1755
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1793..1811)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1879..1983
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1984..2052,2053..2844)
                     /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(2202..2221)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3015..3603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3504..3523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3729..3751
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3748..3764
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3748..3764
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3784..3907
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3948..4253)
                     /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(4050..4069)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4573..5253)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5168..5187
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5307..5313,5314..5331,5332..5337))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5362..5485)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5493..5510)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5493..5509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5502..5524)
                     /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 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
      841 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
      901 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
      961 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     1021 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     1081 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     1141 ccgaggagca ggactgagct agccggccat acaagtaatc cggatgatcg acgccaacgt
     1201 cgttgaattt tcaaatttta aatactgaat atttgttttt tttcctatta tttatttatt
     1261 ctctttgtgt tttttttctt gctttctaaa aaattaattc aatccaaatc taaacatttt
     1321 tttttctctt tccgtctccc aattcgtatt ccgctcctct catctgaaca caatgtgcaa
     1381 gtttatttat cttctcgctt tcatttcatt aggacgtggg gggaattggt ggaaggggga
     1441 aacacacaaa aggatgatgg aaatgaaata aggacacaca atatgcaaca acattcaatt
     1501 cagaaatatg gaggaaggtt taaaagaaaa cataaaaata tatagaggag gaaggaaaac
     1561 tagttctaga gcggccgcca ccgcggtgga gctcccatat ggtgcactct cagtgcaatc
     1621 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     1681 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1741 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     1801 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1861 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1921 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1981 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2041 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2101 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2161 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2221 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2281 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2341 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2401 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2461 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2521 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2581 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2641 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2701 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2761 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2821 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2881 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2941 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3001 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3061 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3121 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggtct tttgctggcc tttgctcaca
     3661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3721 ctgataccag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     3781 tatcaacttt gtatagaaaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
     3841 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     3901 cactatggtc gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag
     3961 aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct
     4021 tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca
     4081 tccccgatat gcaccaccgg gtaaagttca cgggggactt tatctgacag cagacgtgca
     4141 ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc
     4201 acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca
     4261 gcccctgttc tcgtcggcaa aagagccgtt catttcaata aaccgggcga cctcagccat
     4321 cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat
     4381 ggttgtgctt accgaaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg
     4441 tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc
     4501 gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact
     4561 gttatctggc ttttagtaag ccggatcctc tagattacgc cccgcctgcc actcatcgca
     4621 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     4681 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     4741 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     4801 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     4861 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     4921 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     4981 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     5041 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     5101 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     5161 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     5221 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     5281 cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc
     5341 tggggtgcct aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta
     5401 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     5461 cgttcaactt tattatacat agttgataat tcactggccg tcgttttaca acgtcgtgac
     5521 tgggaaaacc ctggcgttac ccaacttaa
//
