LOCUS       sequence-12388-e        5969 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 5969)
  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 5969)
  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: pBCN24-R4R3
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             783..1577
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             783..1577
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(837..856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1447..1466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2153..2175
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2213..2231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2299..2403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2404..2472,2473..3264)
                     /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(2622..2641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3435..4023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3924..3943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4149..4171
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4168..4184
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4168..4184
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4204..4327
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4368..4673)
                     /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(4470..4489)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4993..5673)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5588..5607
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5727..5733,5734..5751,5752..5757))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5782..5905)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5913..5930)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5913..5929)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5922..5944)
                     /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 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
      841 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
      901 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
      961 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     1021 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     1081 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     1141 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     1201 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     1261 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     1321 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     1381 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     1441 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     1501 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     1561 ttgacgagtt cttctgagct agccggccat acaagtaatc cggatgatcg acgccaacgt
     1621 cgttgaattt tcaaatttta aatactgaat atttgttttt tttcctatta tttatttatt
     1681 ctctttgtgt tttttttctt gctttctaaa aaattaattc aatccaaatc taaacatttt
     1741 tttttctctt tccgtctccc aattcgtatt ccgctcctct catctgaaca caatgtgcaa
     1801 gtttatttat cttctcgctt tcatttcatt aggacgtggg gggaattggt ggaaggggga
     1861 aacacacaaa aggatgatgg aaatgaaata aggacacaca atatgcaaca acattcaatt
     1921 cagaaatatg gaggaaggtt taaaagaaaa cataaaaata tatagaggag gaaggaaaac
     1981 tagttctaga gcggccgcca ccgcggtgga gctcccatat ggtgcactct cagtgcaatc
     2041 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2101 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2161 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2221 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2641 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2881 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2941 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3541 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggtct tttgctggcc tttgctcaca
     4081 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4141 ctgataccag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     4201 tatcaacttt gtatagaaaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
     4261 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     4321 cactatggtc gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag
     4381 aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct
     4441 tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca
     4501 tccccgatat gcaccaccgg gtaaagttca cgggggactt tatctgacag cagacgtgca
     4561 ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc
     4621 acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca
     4681 gcccctgttc tcgtcggcaa aagagccgtt catttcaata aaccgggcga cctcagccat
     4741 cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat
     4801 ggttgtgctt accgaaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg
     4861 tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc
     4921 gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact
     4981 gttatctggc ttttagtaag ccggatcctc tagattacgc cccgcctgcc actcatcgca
     5041 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     5101 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     5161 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     5221 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     5281 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     5341 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     5401 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     5461 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     5521 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     5581 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     5641 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     5701 cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc
     5761 tggggtgcct aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta
     5821 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     5881 cgttcaactt tattatacat agttgataat tcactggccg tcgttttaca acgtcgtgac
     5941 tgggaaaacc ctggcgttac ccaacttaa
//
