LOCUS       sequence-12433-e        4480 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 4480)
  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 4480)
  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: pbacNeoR
FEATURES             Location/Qualifiers
     source          1..4480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        158..262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             263..1057
                     /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             263..1057
                     /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(317..336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     927..946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2176..2195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2359..2463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2464..2532,2533..3324)
                     /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(2682..2701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3495..4083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3984..4003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4237..4254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4371..4392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4407..4412,4413..4430,4431..4437)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4445..4461
                     /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     4450..4472
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4469..4480,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4469..4480,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga cgagaaaata ccgcatcagg
       61 cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      121 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      241 ataatattga aaaaggaaga gtatgattga acaagatgga ttgcacgcag gttctccggc
      301 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
      361 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
      421 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
      481 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
      541 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
      601 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
      661 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
      721 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
      781 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
      841 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
      901 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
      961 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     1021 catcgccttc tatcgccttc ttgacgagtt cttctgactg tcagaccaag tttactcata
     1081 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1141 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1201 ccccgtagag gtcggtacca tggtattgat atctgagctc cgcatcggcc gctgtcatca
     1261 gatcgccatc tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta
     1321 catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt
     1381 aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca
     1441 aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat
     1501 aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc
     1561 tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     1621 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     1681 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     1741 gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga
     1801 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     1861 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     1921 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     1981 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2221 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     2281 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2341 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2401 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2461 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2521 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2581 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2641 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2701 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2761 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2821 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2881 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2941 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3001 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3061 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3121 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3181 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3241 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3301 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3361 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3421 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3481 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3541 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3601 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3661 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3721 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3781 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3841 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     3901 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3961 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4021 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4081 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4141 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4201 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4261 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4321 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4381 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4441 ggaattgtga gcggataaca atttcacaca ggaaacagct
//
