LOCUS       sequence-356619-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmYTHDFa_546-700.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6516)
  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"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmYTHDFa_546-700_AC
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2575..2596)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2836..2855)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3183..3204
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3767..3808
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3818..3835
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3858..3875)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3908..4042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3932..3951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3986..4005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4643..5231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4723..4742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5402..6193,6194..6262))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6025..6044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6263..6367)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6435..6453
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtaccatca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2581 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2641 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2701 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2761 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2821 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2881 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2941 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3001 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3061 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3121 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3181 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3241 tacaagccaa gcttgaattc tgcagatatc ccgccaatgc atgggcccga tggcaacaac
     3301 cacgcgcctg ccccactcgc ccgcagcttc aatcgccagg cggacgacaa ggacagggat
     3361 cgcgacggac gcggcggcat gtcacaaaag cacttcaatt ctggaggagg agctggtttc
     3421 cgtaatccag gtcatagggg tggcgccggc agtggcggat cacacaacaa ttacagcgag
     3481 tatcgtcgag gcttcgacat tcataaatca agcagcgact atcagttcaa ggaccgcgat
     3541 ggcccggact ttgacagaga aaatgacttt gggtcccact acgggtccaa taaccgaaaa
     3601 aatgaatata aacacaataa tacgaacaag gccaggttaa aaacccgaga tcgtgacttt
     3661 agtactgaac aaattaaaat cggcgtgcgc tttaaggaca cggacaagga taaaatgcga
     3721 agcaatgaat attcataact cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     3781 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     3841 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     3901 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3961 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4021 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4081 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     4141 ttatgatcag tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     4201 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     4261 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     4321 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     4381 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     4441 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca
     4501 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca
     4561 ggcaggtaga tgacgaccat cagggacagc ttcaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5701 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6001 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6361 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6421 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttaattctat
     6481 attctaaaaa cacaaatgat acttctaaaa aaaaat
//
