LOCUS       sequence-8947-ex        4029 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 4029)
  AUTHORS   Zhang Y, Nash L, Fisher AL.
  TITLE     A simplified, robust, and streamlined procedure for the production 
            of C. elegans transgenes via recombineering
  JOURNAL   BMC Dev Biol. 2008 Dec 30;8(1):119.
  PUBMED    19116030
REFERENCE   2  (bases 1 to 4029)
  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"; journalName: "BMC Dev 
            Biol."; date: "2008-12-30"; volume: "8(1)"; pages: "119"
COMMENT     Sequence Label: pMOD4 GFP
FEATURES             Location/Qualifiers
     source          1..4029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..60
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(91..119)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     867..887
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    2043..2061
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(2200..2219)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2319..2341
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2379..2397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2465..2569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2570..2638,2639..3430)
                     /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(2788..2807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3888..3905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3971..3989
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
ORIGIN
        1 aagcttgcat gcctgcagcc cgggggatcc aaaatggatt acaaggacga tgacgataag
       61 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      121 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      181 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg ggtaagttta
      241 aacatatata tactaactaa ccctgattat ttaaattttc agccaacact tgtcactact
      301 ttctgttatg gtgttcaatg cttctcgaga tacccagatc atatgaaaca gcatgacttt
      361 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
      421 gggaactaca agacacgtaa gtttaaacag ttcggtacta actaaccata catatttaaa
      481 ttttcaggtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
      541 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
      601 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt tgtaagttta
      661 aacatgattt tactaactaa ctaatctgat ttaaattttc agaacttcaa aattagacac
      721 aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc
      781 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa
      841 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt
      901 acacatggca tggacgaact atacaaaagc ccacaagctt tgatgggcac aaatggagga
      961 gtcattgcgg aacagtcaat ggaaatcgaa acaaatgaaa acccagataa ggtagaggag
     1021 cccgtcgtga gaaggaaacg agtaacccgc aggcgccaca ggcggataca cagcaagaat
     1081 aactgcttga cacctccaaa cagcgatgac gaccctcaaa tgagcacacc agatgatcca
     1141 gtcatccaca gtccaccatc aattggagca gctcctggta tgaatgggta tcatggatca
     1201 ggagtgaaac ttgaagaaag ttctggtgct tgtggctctc cggatgatgg cctgttagac
     1261 tccagcgaag agagtcgacg acgtcagaaa acatgtagag tatgtggaga tcatgcgacg
     1321 ggctataatt tcaacgttat cacatgcgaa tcttgtaaag cattcttcag gagaaatgcg
     1381 ttacgtccca aggagttcaa atgtccatat tctgaagatt gcgaaattaa ttctgtaagc
     1441 agacggtttt gtcagaaatg tcgactgagg aaatgcttca cagttggtat gaaaaaagag
     1501 tggattttaa acgaagaaca actgcggcgg aggaaaaact ctcgtttgaa caatacggga
     1561 acgtgtaata aaagatccca gccagggaat cagcagtcac cacaaggtcc taatcaacag
     1621 ccacatttat caccgcatca tccaggagtt gcaatatatc caccacaacc acaacgccca
     1681 ctaacaatca atccaatgga caatcaaatg atgcatcata tgcaagctaa cagaccaaat
     1741 gccatgccac aacttatcag tccaccagga gcacaaccgt atccattgac atccccagtc
     1801 ggctctagtg catctgactc tccaccaaac cgaagtttaa caatgatgca caacggcgaa
     1861 aaaagtccag atggctatga tccaaacatt atggctcatc gtgccccgcc gccatccttc
     1921 aacaataggc ctaaaatgga tagtggacag gttgttcttt ccacggaaga atataaacaa
     1981 ctcttgtctc gaattcgagc caatatgcga gaacacccga gaaaattcat cgatgatggt
     2041 tgagatgtgt ataagagaca gctgtcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2101 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     2161 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     2221 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2281 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2341 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     2401 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2461 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2521 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2581 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2641 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2701 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2761 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2821 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2881 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2941 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3001 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3061 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3121 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3181 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3241 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3301 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3361 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3421 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3481 tttttaattt gtgcatccga agatcagcag ttcaacctgt tgatagtacg tactaagctc
     3541 tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt ttaacgaact
     3601 aaaccctcat ggctaacgta ctaagctctc atggctaacg tactaagctc tcatggctaa
     3661 cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct
     3721 ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa gctagctttt
     3781 aaggtttcac catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3901 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3961 attaagacag ctgtctctta tacacatctc aaccctgaag ctcttgttgg ctagtgcgta
     4021 gtcgttggc
//
