LOCUS       sequence-46683-e        4275 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 4275)
  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 4275)
  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
FEATURES             Location/Qualifiers
     source          1..4275
                     /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"
     primer_bind     complement(1849..1871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1971..1990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2154..2258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2259..2327,2328..3119)
                     /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(2477..2496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3290..3878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3779..3798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4032..4049
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4166..4187
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4202..4207,4208..4225,4226..4232)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4240..4256
                     /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     4245..4267
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4264..4275,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4264..4275,1..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 gcgtcgacgg taccatggta
     1021 ttgatatctg agctccgcat cggccgctgt catcagatcg ccatctcgcg cccgtgcctc
     1081 tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca
     1141 ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt
     1201 taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa
     1261 agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac
     1321 aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata
     1381 gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt
     1441 ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt
     1501 ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt
     1561 ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct
     1621 aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag
     1681 gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt
     1741 ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg
     1801 gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2041 aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg
     2101 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2161 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2221 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2281 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2341 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2401 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2461 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2521 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2581 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2641 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2701 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2761 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2821 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2881 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2941 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3001 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3061 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3121 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3181 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3241 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3301 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3361 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3421 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3481 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3541 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3601 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3661 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3721 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3781 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3841 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3901 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3961 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4021 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4081 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4141 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4201 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4261 acacaggaaa cagct
//
