LOCUS       sequence-288973-        8675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot4_187-1051.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8675)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8675)
  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-DmNot4_187-1051_I
FEATURES             Location/Qualifiers
     source          1..8675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /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(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5960..6001
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6011..6028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6051..6068)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6101..6235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6125..6144)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6179..6198
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6857..7445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6937..6956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7616..8407,8408..8476))
                     /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     8239..8258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8477..8581)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8649..8667
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttgaattctg cagatatcca gcacagtggc ggccgctcga ggatgaccag
     3361 cttggggaca accaaatact gtagccactt catgaagaac cagcagtgtc ccaagagcga
     3421 ctgcatgtac ttacatgaat tgggcgatcc cgaggccagt ttcacaaagg aggaaatgca
     3481 tcagggaaag cacctggagt acgagaagcg cctgcacgat actctaattg cgtcattggg
     3541 accgaatgca acaggtattg taaatgtcgc gattccatcg tcatcgtcag cgtcatcctc
     3601 gtcctctgga tcgggtacaa atggttcaag tgcatcaggt aatgcccagc aaaaggaggc
     3661 ctggccaagc ttatcggtct cgcccatcaa tggcaaagag gcggctgcca ctgcgaccag
     3721 ttccagtggg aaaagtaaac gggagaagct gcgcaacgag aagaggcacg agaagaacaa
     3781 ggcgaagaat aagaacggca gcaatacaaa cgccaatgcc tcaaacaagg agaactatgt
     3841 tcccgaaacg agaagcagca caagtactga gacattcgca gaggctacgg cggatgcacc
     3901 ggcttccacc aaggcagaac cgccgcaagc ctctagcaat agaacgaggg cggatcgtgg
     3961 aaaagatcgg accacggcta gtgcaaagga gcagaagaag ggcaaggaaa ctgctccagc
     4021 acctgcagca agtaaaccgg cggagcgggt tgaaacaagc gagagtacaa taagacaaaa
     4081 gaaggcggaa gtaaccgaaa gctgtgaaga taacttacca caaaagagat tagcgggaac
     4141 aaacgttcaa agatctgtga gctcttgtag cgaaaatagc gaaggacacg tctctgagag
     4201 tagcttaagt gagaagagtt taactggtga ttatgtggag gaaaagtgca atagtgtgaa
     4261 ttcggaaagc cagcaagaaa gtgtaaagtt tcaggaggag ctcgaaaaga gcaacgaggc
     4321 aattgtcgag gccgaaacga ttctgccaac agctgagtcc tctgaggaca tcagtcccgc
     4381 tgcagttgcg ccaggcaatg gagaagacga aggatgctta cccgttgttg atccagtcga
     4441 accaccgagt ctggcggata atggaagcag agtaacagat gcgctatcta agctgaacat
     4501 tttcgatgac acgcctagct ttttcacatc gccatcattc cagcaagcac ccatattaaa
     4561 gaataagcta gatttggaaa tgcgacagtc tcatttacct gacttggtca acgacattga
     4621 tggaatccaa aaggcatcca atacaaacga gtgggaggaa gcctttaaaa atgtgatgat
     4681 gggcaacacg cagcacatgg aggagcaact gttgcagcag caacatttgc agcagcatca
     4741 gaatttgcgc caccagctgg tgttgcaaca ggaggagttc cttcgcatgc aagaattaca
     4801 gaaacgcaac aactttgcga cgcaaatcaa cggtcctgcg aatgatttcc ttagagctta
     4861 tgagctgaga gctcaagcca atgcaatcat tcagcaacaa ttattacaac agcacgcagg
     4921 cgaaaatcta tttggcggca acatgtcgaa gttcttcgat ttccataaaa gtcagccgca
     4981 gtcacaccat cagtatctaa atggacatcc gcctcaaatc aatgggaatg gtgctgtgcc
     5041 ggagccgcaa agagtggcgg cctctttgga aagcaatcga ctgaattcgc cttttgtcga
     5101 aaatggactt attaactcgc agcagcagca gcagcagcag caacagcaac aaaagcagag
     5161 aatgatggga atgtatgaat ttatgcctcc aaacacgcaa tctcagcaga atcggtttac
     5221 acagaactcg atagtcgacg atgacttagg attcgacccg ttcgttgaga cccaaaaggg
     5281 acttgctgaa cttatggaaa atgaggtggt ccaaaaacag agtattaata atgagaaccc
     5341 cttgccgaag ttgccgccac agcctcaagt tccaccccat ccgcaattgg tggacaacct
     5401 gcagcgagct cgcatgccgc cgccaggctt caatcacgtc aacacattgg gcttgggcgg
     5461 tgcatccaga ctgcagctca ccagcaaaat aatgcccttc atgaacatgc ccgttaatgg
     5521 ggtgggcaac agcggtgccc agggacagca ccaaatgcca atgggtgtca actggaatgc
     5581 gccgatgggc atgcaccaga acccgggaca gcccgtgggt gattcgcaat tgcagcatcc
     5641 aatggctcac aacaaggttt acaacaacag tgattggact tcaatggatc cggctatact
     5701 ttcgtttaga cagtattctt cttttccaca aaaccaaatt cccccacatc ctcagcagca
     5761 acaggattta tttttgcagc atttggctca acagcaaaat tcccagagtg gtggtttcaa
     5821 caaccaggca cagcaaatgc ttcctatggg gatgcccaat agtttgctga acggacaaca
     5881 aacgcagccg ccacaggtca atgccaatgt tcagggcatg cttgagtttt taaaaagccg
     5941 tcaattcgtt tagttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     6001 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     6061 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     6121 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     6181 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     6241 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     6301 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgaccg atgcccttga
     6361 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     6421 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     6481 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     6541 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     6601 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     6661 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     6721 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     6781 agcttcaagg atcgctcgcg gctcttacca gccagcaaaa ggccaggaac cgtaaaaagg
     6841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7021 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     7081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7261 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7921 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8221 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8581 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     8641 cctataaaaa taggcgtatc acgaggccct ttcgt
//
