LOCUS       sequence-486273-        2604 bp    DNA     circular SYN 24-SEP-2025
DEFINITION  Encodes a fragment of eGFP for generating dsRNA for use as a control
            in RNAi experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2604)
  AUTHORS   Deihl EW, Booth CLT, Davies EL
  TITLE     Protocol combining RNA interference and regeneration assays in 
            planarian embryos.
  JOURNAL   bioRxiv [Preprint]. 2025 Jul 22:2025.07.21.666018. doi: 
            10.1101/2025.07.21.666018.
  PUBMED    40777484
REFERENCE   2  (bases 1 to 2604)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Jul 22:2025.07.21.666018. doi: 
            10.1101/2025.07.21.666018."
COMMENT     Sequence Label: pDL1-eGFP
FEATURES             Location/Qualifiers
     source          1..2604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(249..353)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(431..478)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     480..496
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        497..515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        517..535
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(539..558)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     886..907
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(951..969)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(952..969)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(971..989)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(990..1006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(990..1006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1008..1055
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1062..1079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1233..1821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1313..1332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1992..2604,1..179,180..248))
                     /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"
ORIGIN
        1 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
       61 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      361 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      421 ccgcatcagg caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctaggg
      481 taaaacgacg gccagttaat acgactcact atagggaatt aaccctcact aaagggagga
      541 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      601 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      661 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
      721 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
      781 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      841 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      901 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt tctatagtgt
      961 cacctaaatc cctatagtga gtcgtattag tcatagctgt ttcctgtcta gcataacccc
     1021 ttggggcctc taaacgggtc ttgaggggtt ttttgcgctt cctcgctcac tgactcgctg
     1081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2281 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2581 cgaccgagtt gctcttgccc ggcg
//
