LOCUS       sequence-12436-e        8253 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 8253)
  AUTHORS   Giordano-Santini R, Milstein S, Svrzikapa N, Tu D, Johnsen R, 
            Baillie D, Vidal M, Dupuy D
  TITLE     An antibiotic selection marker for nematode transgenesis.
  JOURNAL   Nat Methods. 2010 Sep;7(9):721-3. doi: 10.1038/nmeth.1494. Epub 2010
            Aug 22.
  PUBMED    20729841
REFERENCE   2  (bases 1 to 8253)
  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.1494"; journalName: "Nat Methods"; date: "2010-09"; 
            volume: "7"; issue: "9"; pages: "721-3"
COMMENT     Sequence Label: pRG5273Neo
FEATURES             Location/Qualifiers
     source          1..8253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             795..1589
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(849..868)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1459..1478
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2290..2306
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2291..2307
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3971..3989)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4010..4026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4010..4026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4023..4045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4034..4050
                     /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)."
     promoter        complement(join(4058..4064,4065..4082,4083..4088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4241..4258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4412..5000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4492..4511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5171..5962,5963..6031))
                     /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     5794..5813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6032..6136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6162..6617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6299..6320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6511..6530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6744..6766
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6758..6775
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6759..6775
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6782..6801
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6782..6800
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atttctaaat tttagtttaa taaaaaaatt ttaaaataat acgagttttg agactaactt
       61 tcaataaaaa ccaaaaaatt gcttcaaaaa agagctcgcg ccgatttacg ataaacacag
      121 cgcgctacga cactccgaaa accgcgccgc accattacat tctccgcccc accgcacggt
      181 ttgcggaagg cgatgaaatt gctgggccta aaattttaac cctttttttc ccgtttttca
      241 actgttttcc cgattttttc gatatttttt tcaggttttt agttagaaaa gtatataaaa
      301 attgcattat ttgtagtttt cgctcgtaat ttgtgataga aaacggccaa tttgcctaga
      361 tttgtgattc taccaagtgg aatttttggg aaaatatcaa tattccacga tttttattca
      421 aatcataacg tttcgaaatt atcagcttaa ataaaactaa aataaataat aaaatactaa
      481 aaaccctgaa tttctcggtt ctcggttcag cttgagttcg ttctatacac gttatcgatt
      541 tttgtcattt cgtttaatca tgggttttga ttcattttaa atatgaaatt tgatttattc
      601 ctcgtttcga agttggtttg tcttgtccgg ccggtaaaca aatgttatta aaaaaaataa
      661 taaatttcga aatttttcgt aatgttccgc aatcgtgttt ttccgtttat ctcgtcttcc
      721 cttccttcaa atatactatt tattttctca attcacatat tttcagccag ccctgctcaa
      781 caagtggaat aaaaatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
      841 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
      901 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
      961 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     1021 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     1081 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     1141 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     1201 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     1261 atgatctgga cgaagagcat caggggctcg caccagccga actgttcgcc aggctcaagg
     1321 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     1381 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     1441 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     1501 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     1561 tctatcgcct tcttgacgag ttcttctgag tccaattact cttcaacatc cctacatgct
     1621 ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc
     1681 tttgtttttt agcttctttt aagccacctc taacaatgaa attgtgtaga ttcaaaaata
     1741 gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat
     1801 tctatcccaa aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa
     1861 atttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt
     1921 cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca
     1981 tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat
     2041 gaaattaatt ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga
     2101 ggacggcgtt tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg
     2161 aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg
     2221 agtagtgtct atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat
     2281 aactgtttct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggt
     2341 agattacatc aagctcaaga atttttagtt taacctcgtt tttatgtcta gggattgttt
     2401 tcctgagcgg attgtatcca atgcacccac agctaaatct gaaaattaga tttcagtgaa
     2461 agttttgaag gattcatgac gtggacccac ttgaaagtca acctctccat atgttacatg
     2521 tcctttcaag taaattacag acagttcaaa aaagcaaaga atatttagga aaacatgagt
     2581 gggccgaaga aatacgacac cctcaatccg aattccgtca taaatcaaaa gttcttgaaa
     2641 ttttgcaaag tactctaaca ttgttattac gcgccgcggc gtagcttttt ttaaaagttt
     2701 cagctggtag tagaaactga atgagagaac aaattaaact ccgaattatg gaaatagaca
     2761 atagactccg ccgccctttt gatagtgcta ctatgcaacg tgatttgcac atataaattc
     2821 atgtagccaa gttttttgat ctcttaattg caagtttcaa aattttcatc gcaactacac
     2881 gtctttgtct tgtgtcacga gtagcgtcac aaactcccat ctcaaacatt actcattgaa
     2941 aacgcaatca gttaagcact agattaaccc ttaatctgat ccacaacctg aaagtgatct
     3001 tcaaataaaa attcaaaagg aggtggctcg attgagcact tgttttatgt catatgggat
     3061 tttagtaagc aaagttctta ttttgctttt atttggcgtt tttttttggg gattttgaaa
     3121 aaaaaaacgt tttactaatt attgcaatat ttacctaggg atgagaattt tcaaacttcc
     3181 attttgtttg aaacataagt attaaaatat tattagaagg ttctttcttt ctgcagctga
     3241 cctgtttgag acacattttt ttattttatt tataaaaatt gatgattctt gttgccgttt
     3301 ccacaatgtc ctcattcagt aaacttgtga gattacaatt ttatttcgtt tgtgtctcct
     3361 gttttttaaa attcaattat tttttaaata cataccatat gccgtttctt ttcaatattt
     3421 acatctgtct ggtaaatgat gtgtattaat ttattagtta gtcatttgtt ttctttttca
     3481 ttcaactatt ttcattactc tcccacgttt ttccaaccaa tcgcgtcgac ggaaatgtaa
     3541 aatgtgagat attgtgcggc ttcttctttt aaattcgcca taatttattt cactctttcg
     3601 gtcacacgat tttttttcca caattggcac cttaaatttg acgaggctaa gaaaccaaag
     3661 gtcctcctgc tttccggtga ctcgcttata gtatctgagt gctagtaaac aaagttcttg
     3721 gaacttgtgt aaagcttaat gaaggatcca gaatatgtga ttactgcctg cttctaatta
     3781 catagaggat atgatgcgca aatttttgaa agagcgatag ttcagtttag tttcggctgt
     3841 gagtggcagc gcatctttta ttgcttcgat tgttttgttc ctcatctcat ttcttacaaa
     3901 gtccagatat ttagagtgac gatgcccatc atccgcggcc gccaccgcgg tggagctcca
     3961 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     4021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     6181 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     6241 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     6301 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6361 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6421 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6481 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     6541 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6601 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6661 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6721 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     6781 gtaatacgac tcactatagg gcgaattggg taccgtatta taattccaaa aacacagttt
     6841 atttgaaata tgcgcataaa accttcgtta gcatagtcca tgacggaaca taatgcgcga
     6901 atcgcgaaaa taattgtttg caatgcattg tgcctcattt cgttatttac atatgtgtgc
     6961 atcatgtcct tggttcctgc cgattacatt gtaaatgaac tgtgaaagta cagtgtactt
     7021 tgtcctagaa ggacacgttc tttctcatat tcgtggattt attcgtgaaa ttgaaaactg
     7081 acattcacat atgttttttc atgagaatag aagttgcaag gttccctgga tattcagtaa
     7141 ttacattttt accaatgtca cacactaagg ggattaagaa ggtcggtatg agggtgggtt
     7201 gggtggcaaa cgcagtacga ttaatctatg taccttttcg ctttctcatc aacaaaatgg
     7261 gcattctctt cgaatttaca taatttcaaa attacttttg tttttaatat aatccttcga
     7321 atacattaca ccattttgca ataagtcata gatcatcttt tttgagaaac ctcgcttttc
     7381 gttaaaacta tacatttaaa caaaactctc tttaatagag agcaataatg aggtgacgtt
     7441 taaatgagtt ttttgtatta atttttagcg gaaaggagaa ggaacttgtg aaaaaaaaat
     7501 tgaaaaggaa cgggtttagt ctgccagaat tcctgagtat tgtcagtctg actttatatt
     7561 ttgagctcag gttcatagtt tttggtttct aaattagaac tagcacaggg cagcttacta
     7621 catattagtt ttgaactttt attttaatga ttatgtatta tattcccaca cttttaaata
     7681 aaaattttat agttataaaa atgaaagcgt gaggaatatt tatgtatttt taggcgggga
     7741 tataatacac aagtatattt ttcaaaaata tattttcatc ttaatcttta aaataggaag
     7801 tgtaagcaat tttatgatct gacaactaac aggattcacg cagcactatc aaaaccttca
     7861 cagggcgcaa aacagctgcg atacaaaccg tatttcttct attagtatgg ttgcaatact
     7921 atttttcgaa ggtcttcccg tatgcaatac taatagggag tccaagtcta ataaggaggc
     7981 cataatgaag aaatacggtg tatagtaccg cgcgaaaccc ggtagacgtc gggccacgat
     8041 agtaattacc ctttcaactt taaagacttg tcagaaaatg tggatttaac tttaacagca
     8101 atatcagtaa cgtccctcag attttcaatt ttcaataatt tttattcctg tctatttttc
     8161 gccttttcta aataggaata gttaatgaaa atgttcctgc ttaaaattgt atgataatca
     8221 ttattttgcc tcgtcactca attcgacggg ccc
//
