LOCUS       sequence-289605-        8782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmBicaudalC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8782)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8782)
  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-DmBicaudalC_P
FEATURES             Location/Qualifiers
     source          1..8782
                     /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      4983..4992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6067..6108
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6118..6135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6158..6175)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6208..6342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6232..6251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6286..6305
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6964..7552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7044..7063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7723..8514,8515..8583))
                     /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     8346..8365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8584..8688)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8756..8774
                     /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 ttgatgttgt cctgtgcctc tttcaataaa ctgatgtatc cgtcggccgc
     3361 agatgttgcc aaaccgccaa tggtgggctt ggaggtggaa gctggatcca ttggatccct
     3421 gtccagtctt cacgcccttc catcgaccac ctcggtgggc agtggagccc ccagcgagac
     3481 gcagagcgag atctcctccg tggatagcga ttggagtgac atacgcgcca tcgccatgaa
     3541 gttgggcgtt caaaacccag acgatctcca cacggaacgt ttcaaggtgg atcgacagaa
     3601 attggagcag ttaataaaag cggagagctc cattgaggga atgaatggag ctgaatattt
     3661 tttccatgac atcatgaaca ctacggatac ttatgtgagc tggccttgca gactcaaaat
     3721 tggtgccaag agcaagaagg atccccatgt gcgaattgtg ggaaaggtgg atcaggttca
     3781 gcgggcaaag gagcgcatcc ttagcagtct tgactccagg ggcactcgag tgatcatgaa
     3841 aatggacgtc agctacacgg atcactcgta catcatcgga cgaggtggca acaacatcaa
     3901 gcgcataatg gacgacacgc atacccatat ccacttcccc gactcgaatc gctccaatcc
     3961 cacggagaaa tccaaccagg tgtcgctgtg cggcagccta gagggcgtgg agcgagcccg
     4021 ggctctcgtt cggctctcca caccgctgct catctccttc gagatgcccg taatgggacc
     4081 taacaagccg cagcccgacc acgagactcc ctacatcaag atgatcgaga ccaagttcaa
     4141 tgttcaggtc atattctcca cgcgtccaaa attgcatacc tcgctggttt tggtcaaggg
     4201 atccgagaag gaatcggcgc aggtccgaga tgccacgcag ctgctgatca acttcgcctg
     4261 tgagagtatt gcgagccaaa tcctggtgaa tgtccagatg gagacctcgc cgcagcatca
     4321 cgaaatcgtc aagggcaaga acaacgtgaa tctgctgagc attatggagc gaacccagac
     4381 caagatcata ttccccgatc tgagcgacat gaacgtgaag cccctgaaga agtcccaggt
     4441 cacgatcagc ggacgcatcg acgacgtcta cttggcgcga caacaattgc ttggcaattt
     4501 gcccgtagca ttgatttttg actttccgga caaccacaac gatgcatccg aaataatgag
     4561 cctcaacacc aagtacggcg tctacatcac gttgcgccag aaacagcggc agagcaccct
     4621 ggccattgtg gtcaagggag tggagaagtt tatagacaaa atctacgagg cccgccagga
     4681 gatcctgcgc ctggccacac cattcgtgaa gcccgagatc cccgactact acttcatgcc
     4741 aaaggacaaa gaccttaatc tcgcctaccg cacccagcta actgctctgc tggctgggta
     4801 tgtggacagc ccgaagactc catcgctgtt gccgccctcc ttggctggcc aactgacgcc
     4861 ctatgccaac aacaatcacc tgttgttgaa cgccaatggc ttggccacgc ccactggggt
     4921 gtgtgcgccc acccaaaaat atatgcaatt gcacaacagc ttccagcaga ctcagaatcg
     4981 ttccatggtg gctggtgggc agagcaacaa tggcaactat ctgcaggttc ccggagcggt
     5041 ggcgccgcct ctcaaaccgc cgactgtttc gccacggaac agctgcagtc agaataccag
     5101 tggttaccag agctttagca gcagcaccac ctccctggag cagtcgtatc cgccgtatgc
     5161 ccaattgccg ggcaccgtgt cgtccacttc atcctccacc gcgggaagcc aaaatcgggc
     5221 gcattattcg ccagattcca cgtacggcag tgaaggtggt ggcgttggcg gcggaggtgg
     5281 tggtggggcg cgtctgggac gacgccttag cgatggagtg ctgctggggc tgggtaattc
     5341 caacggtgga ggaggtaact ccggaggagc ccacctcctg cccggcagtg ccgagagcta
     5401 tcggagtttg cactatgacc ttggaggaaa caagcactcc ggccaccgtg cctttgactt
     5461 cgacatgaaa cgggccctgg gctacaaggc catggagcgc accccggtgg caggggagct
     5521 gcgaacaccc acgaccgctt ggatgggaat gggcttgagc agcacctcgc ccgcccctgc
     5581 cccgctggag aacggggaaa atggggctgc tggtggagga gcatctagtg ggtggcggct
     5641 gccaccagga ctgggttccc cttacggact gagtgccacc accggactcc tggacgccac
     5701 tccagtcaac cgacggatgc agctagccaa gcacaaggac atccagactc tgctcaccag
     5761 tttgggtctg gagcactaca tcaaaatatt cgttctaaac gaaatcgacc tggaggtgtt
     5821 caccaccctg accgaggaaa acctgatgga gctgggtatc gcggcgttcg gagctcgcaa
     5881 gaagctgctg acggccattc acacgctgct ggccaacgag gcggcctgca gcacaatgcc
     5941 cagcagcagc tcctcgcaga actcctcgtc gccgcgattc tccggaagtg cggccccagg
     6001 cgcagagcgc cgtccctcca accagtggtg agcggccgct cgagtctaga gggcccgcgg
     6061 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat
     6121 catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc
     6181 tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat tgatgagttt
     6241 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     6301 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     6361 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     6421 tacaaatgtg gtatggctga ttatgatcag tcgaccgatg cccttgagag ccttcaaccc
     6481 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     6541 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     6601 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     6661 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     6721 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     6781 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     6841 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     6901 gctcgcggct cttaccagcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6961 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7021 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7081 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7141 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7201 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7261 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7321 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7381 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7441 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7501 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7561 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7621 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7681 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7741 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7801 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7861 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7921 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7981 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     8041 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8101 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8161 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8221 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8281 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     8341 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8401 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8461 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8521 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8581 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8641 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8701 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8761 gcgtatcacg aggccctttc gt
//
