LOCUS       sequence-288795-        8918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGe1_1-943.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8918)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8918)
  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-DmGe1_1-943-V5His6_G
FEATURES             Location/Qualifiers
     source          1..8918
                     /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"
     regulatory      5163..5172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6203..6244
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6254..6271
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6294..6311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6344..6478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6368..6387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6422..6441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7100..7688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7180..7199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7859..8650,8651..8719))
                     /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     8482..8501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8720..8824)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8892..8910
                     /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 ggccgccgca tgttaatcgc
     3361 gctcttcgcg cttgcacact tgccaatctt tcgtaagacg cctttgtcag cctcctcctc
     3421 cccgccgccg tctgtccacc ggtcgccaag gtgtggcaaa gcaagtaccg cgtaccatcc
     3481 accgccagca gacgcctgca tcagcattac tgcttcagcc attgacgcag catccgggat
     3541 gagcaacagc gccgaacagg acaagccgat tgccgccaac ggcaatgcgg cagagcccca
     3601 ggaaatagaa gtcatccatt tccaagcgca agagaagcag tgttgccgcg ttattagatc
     3661 caataagacc aaggtcctca catccggcgg cgttcacaca cgcggttcct ccaaggtcaa
     3721 gctaaagaac atcgtcgact acaagtggga gcgcaagtac tactatccgg gtcacctggt
     3781 cgctgtccac cgcgatggca agcacctggc gtacgccatt aatgtcaaca acaaagccac
     3841 cggaatggag ggcatggtgc gcgtatgcaa catagccacc agtatgcgag ccttgatcaa
     3901 gggaatgagc ggcgaggtct tagatctgca gtttgcccat acggattgtg agcgtatatt
     3961 agctgtgatc gatgtcagtt cgctgtttgt ctacaaagtc gatcaaatcg aagggaacct
     4021 gctgtgcaat ctagtgctaa aggtggagga ccccatcgcc aactatgtgc cggagtacga
     4081 tatggtctcc tggtgtccct atgtctgcag cagcagcgcc accgttccaa tcaacgacga
     4141 tgacgatgag aatcagctgc tcatttggtc gcgaagctct caatttcaat gctttcaagt
     4201 gaagatgatt gtctctgaac atgggcgcgg caagatccaa ccagcagccc tagaatctgg
     4261 atatctgaag attgaggagg actcgttgat cacgtgcgcc gctttgtccc cggatggcac
     4321 cactgtggct gctgcttgcg ccgatggctt ggttcgtttc taccaaattt atttgttcga
     4381 tgtgcgcaac catcgctgcc tgcacgagtg gaagccacat gatggcaaaa aagtctgctc
     4441 tctctttttt ctcgacaata tcaacaagcc tgtggaagaa tcctactggc agcacgtgat
     4501 cactacgagc gatgccaata ccgaaatcaa gctctggaac tgctcgctat ggaaatgcct
     4561 ccaaactatc aatgtggtgg ccagtccgtc atctttgcag cccagaaact ttatcgcggg
     4621 catagatcgc agtgcgaact acttggtgtt gtcttgcctg gactctctgg ccgtgtatgt
     4681 aatgcagatt ggcagtactg gcggcgcaga ctccgagaat aggagtagcg atagcgaagg
     4741 agaaggctgc gacacttcca aacgcattca gaatgttgcc gaattcaagc tgagttcggg
     4801 gattctctct ttttctattg taaacgctag tatgcgtcgc gttaagaact ccatcgagag
     4861 ctactatcct attgaggagc ccgatgactt cgatgacgat tccaattcca ccagtgccct
     4921 tgtgctgcac atgtttgtgg tgcaggcaaa gagcctgcag gaatgtcaaa taatctacca
     4981 gccatgtgtg gctgaaaaga cggagcggag cagcctaaac agcaagcgat cacagacgcc
     5041 agaagacaat ctgctaatca aagaggagcc ggaatccccc aacagtggaa ctgtgggtgc
     5101 tgttcagctg gatgcattat tcgcaaaatc cgcaaagaga gcttcaacgg gcagttcatc
     5161 tgccatggtg gccgtggcgg cagccgcggc tgcggcacct tcggccatcc tgcaggacgc
     5221 cactaaggag gcggccaagt cagagtcccc gcaactaagc agcgcgtata cgcagcaggt
     5281 taatctcatg actccagatg ccttcagtgc aagcggaacc gcagcggcag cagctgtatt
     5341 tgtgtctacc agcaccacaa cttcaatagg aactgatagc tctaccacca ccagcggtca
     5401 ggatagatcc attgattcag cggttctgca aactatacga atgctggcca cggtaacgtc
     5461 caagacttcc gaaaacccca atgccgaggt actcctcaac ctaatgaaca acaccctaat
     5521 cgaagatcgc gagcaacaaa agctgaagga aaagcttgac gctcgcaaga agttcattgc
     5581 aattgatcgc aatccagagc ggaatgtggc tgagaacttg gccagcggca gttcgagtcc
     5641 gagtcgagag gtgcaggaga taatggccac acaggacgat gccgatgcct acgaagcaga
     5701 gctagagaat ctggacgacg atgatgacga tgaggaagaa gagctggcaa attcatcacc
     5761 gctcccggaa gctgttgacg gaacatggcc cattgtcaag ctgtcatccc actcagctga
     5821 gctccagaat gctgcccaga tcatgtccca ggcggtgcag aacaccaaca atggcaatgt
     5881 gccgcctacg ctaggtggag ggcacaataa taacacaagt gtaggcagca atagcaacaa
     5941 caacacggct acaactctta gcaccagcaa caccagcagc agcaacaacg cgggcggaac
     6001 atgcgttgat tcaagtggaa ctggagagct aaatgccaaa atggaactac tcatagatct
     6061 agtaaaagcc cagtccaaac aaataaacaa attggagaac gaagtcaaca agctacaaaa
     6121 gcagcaagaa gcggcggcgg ccctgcatag caaacaagac acttcattgg agcctaaaag
     6181 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     6241 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     6301 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     6361 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     6421 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     6481 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     6541 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga ccgatgccct
     6601 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     6661 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     6721 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     6781 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     6841 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     6901 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     6961 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     7021 gacagcttca aggatcgctc gcggctctta ccagccagca aaaggccagg aaccgtaaaa
     7081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7261 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     7321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7501 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8161 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8461 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8821 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8881 taacctataa aaataggcgt atcacgaggc cctttcgt
//
