LOCUS       sequence-46682-e        5447 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 5447)
  AUTHORS   Semple JI, Biondini L, Lehner B
  TITLE     Generating transgenic nematodes by bombardment and antibiotic 
            selection.
  JOURNAL   Nat Methods. 2012 Jan 30;9(2):118-9. doi: 10.1038/nmeth.1864.
  PUBMED    22290182
REFERENCE   2  (bases 1 to 5447)
  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"; 
            journalName: "Nat Methods"; date: "2012-01-30- 30"; volume: "9"; 
            issue: "2"; pages: "118-9"
COMMENT     Sequence Label: pBacPuroR-NeoR
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        140..244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             245..844
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(245..264)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     741..761
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     promoter        1125..1229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1230..2024
                     /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             1230..2024
                     /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(1284..1303)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1894..1913
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3021..3043)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3143..3162
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3326..3430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3431..3499,3500..4291)
                     /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(3649..3668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4462..5050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4951..4970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5204..5221
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5338..5359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5374..5379,5380..5397,5398..5404)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5412..5428
                     /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)."
     primer_bind     5417..5439
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5436..5447,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5436..5447,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgagaaaa taccgcatca ggcggcctta agggcctcgt
       61 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      121 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      181 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      241 gagtatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
      301 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
      361 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
      421 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
      481 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
      541 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
      601 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
      661 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
      721 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
      781 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
      841 ctgactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
      901 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
      961 tgagttttcg ttccactgag cgtcagaccc cgtagagcta gcgtcgacga gaaaataccg
     1021 catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1081 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1141 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1201 tgcttcaata atattgaaaa aggaagagta tgattgaaca agatggattg cacgcaggtt
     1261 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     1321 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     1381 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     1441 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     1501 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     1561 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     1621 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     1681 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     1741 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     1801 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     1861 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     1921 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     1981 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgactgtca gaccaagttt
     2041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2161 cgtcagaccc cgtagaggtc ggtaccatgg tattgatatc tgagctccgc atcggccgct
     2221 gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac
     2281 atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaac
     2341 ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt
     2401 agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc
     2461 cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt
     2521 ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc
     2581 atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca
     2641 tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca
     2701 agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc cctattgttt
     2761 gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat gagcacgatg
     2821 caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt agaagttgat
     2881 aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa tacattccca
     2941 atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg tctcgcgcgt
     3001 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3061 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3121 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     3181 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg
     3241 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3301 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3361 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3421 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3481 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3541 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3601 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3661 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3721 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3781 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3841 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3901 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3961 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4021 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4081 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4141 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4201 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4261 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4321 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4381 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4441 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4501 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4561 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4621 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4681 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4741 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4801 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     4861 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4921 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4981 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5041 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5101 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5161 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5221 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5281 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5341 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     5401 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
