LOCUS       sequence-364073-        3040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhanced RNAi clone for C. elegans knockdown of non-codon-optimized 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3040)
  AUTHORS   Palmisano NJ, Azmi MA, Medwig-Kinney TN, Moore FEQ, Rahman R, Zhang 
            W, Adikes RC, Matus DQ
  TITLE     A laboratory module that explores RNA interference and codon 
            optimization through fluorescence microscopy using Caenorhabditis 
            elegans
  JOURNAL   CourseSource, 2022
REFERENCE   2  (bases 1 to 3040)
  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: 
            "CourseSource, 2022"
COMMENT     Sequence Label: T444T-GFP(NCO)
FEATURES             Location/Qualifiers
     source          1..3040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     177..198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        361..465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(466..534,535..1326)
                     /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(684..703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1497..2085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1986..2005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2147..2194)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2195..2213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2229..2942
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2259..2287)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2882..2902
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(2978..2996)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      join(2997..3040,1..4)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tttggccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
       61 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
      121 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
      181 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
      241 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
      301 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
      361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      481 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      541 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      601 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      661 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      721 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      781 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      841 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      901 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      961 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1021 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1081 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1141 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1201 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1261 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1321 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcaaa aaacccctca
     2161 agacccgttt agaggcccca aggggttatg ctagtaatac gactcactat agggagaccg
     2221 gcagatctat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     2281 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     2341 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     2401 caacacttgt cactactttc tgttatggtg ttcaatgctt ctcgagatac ccagatcata
     2461 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     2521 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     2581 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     2641 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
     2701 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     2761 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     2821 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     2881 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     2941 aagtcgacct cgaggggggg cccggtaccc aattcgccct atagtgagtc gtattactag
     3001 cataacccct tggggcctct aaacgggtct tgaggggttt
//
