LOCUS       sequence-8950-ex        4615 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 4615)
  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 4615)
  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 RT-G
FEATURES             Location/Qualifiers
     source          1..4615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..56
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             926..2116
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1278..1297)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    2629..2647
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(2786..2805)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2905..2927
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2965..2983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3051..3155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3156..3224,3225..4016)
                     /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(3374..3393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     4474..4491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4557..4575
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga tggattacaa ggacgatgac gataagatga
       61 gtaaaggaga agaacttttc actggagttg tcccaacatg tcgtcgtaac ctgtagaacg
      121 gagtaacctc ggtgtgcggt tgtatgcctg ctgtggattg ctgctgtgtc ctgcttatcc
      181 acaacatttt gcgcacggtt atgtggacaa aatacctggt tacccaggcc gtgccggcac
      241 gttaaccggg ctgcatccga tgcaagtgtg tcgctgtcga gatatgacgg tgttcacaaa
      301 gttccttaaa ttttactttt ggttacatat tttttctttt tgaaaccaaa tctttatctt
      361 tgtagcactt tcacggtagc gaaacgttag tttgaatgga aagatgcctg cagacacata
      421 aagacaccaa actctcatca atagttccgt aaatttttat tgacagaact tattgacggc
      481 agtggcaggt gtcataaaaa aaaccatgag ggtaataaat aatggcaaca gttaaccagc
      541 tggtacgcaa accacgtgct cgcaaagttg cgaaaagcaa cgtgcctgcg ctggaagcat
      601 gcccgcaaaa acgtggcgta tgtactcgtg tatatactac cactcctaaa aaaccgaact
      661 ccgcgctgcg taaagtatgc cgtgttcgtc tgactaacgg tttcgaagtg acttcctaca
      721 tcggtggtga aggtcacaac ctgcaggagc actccgtgat cctgatccgt ggcggtcgtg
      781 ttaaagacct cccgggtgtt cgttaccaca ccgtacgtgg tgcgcttgac tgctccggcg
      841 ttaaagaccg taagcaggct cgttccaagt atggcgtgaa gcgtcctaag gcttaatggt
      901 tctccaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      961 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     1021 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
     1081 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
     1141 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
     1201 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     1261 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     1321 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     1381 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     1441 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
     1501 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     1561 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     1621 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     1681 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     1741 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     1801 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     1861 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     1921 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc
     1981 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     2041 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     2101 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     2161 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     2221 atatccatcg cgtccgccat ctcgaccaat tctcatgttt gacagcttat catcgaattt
     2281 ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac
     2341 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     2401 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     2461 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatgcttct tgagtttgta
     2521 acagctgctg ggattacaca tggcatggac gaactataca aaagcccaca agctttgaat
     2581 tcgagccaat atgcgagaac acccgagaaa attcatcgat gatggttgag atgtgtataa
     2641 gagacagctg tcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2701 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2761 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2821 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2881 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2941 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3001 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3061 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3121 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3181 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3241 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3301 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3361 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3421 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3481 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3541 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3601 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3661 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3721 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3781 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3841 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3901 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3961 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4021 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttgtgc
     4081 atccgaagat cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta
     4141 ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct
     4201 aacgtactaa gctctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
     4261 atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt
     4321 tttataagaa aaaaaagaat atataaggct tttaaagcta gcttttaagg tttcaccatg
     4381 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4441 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4501 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta agacagctgt
     4561 ctcttataca catctcaacc ctgaagctct tgttggctag tgcgtagtcg ttggc
//
