LOCUS       sequence-288445-        7823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmSmaug_1-583.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7823)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7823)
  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-DmSmaug_1-583-V5His6_C
FEATURES             Location/Qualifiers
     source          1..7823
                     /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             5108..5149
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5159..5176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5199..5216)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5249..5383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5273..5292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5327..5346
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6005..6593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6085..6104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6764..7555,7556..7624))
                     /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     7387..7406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7625..7729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7797..7815
                     /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 cagatatcat gaagtacgca actggaactg acaacgcgat
     3361 gacctccggc atcagcggcc aaaccaataa ctctaacagt gcatccaacg agatgcagcc
     3421 aaccacatca acgcctactg cagcccacaa ggaggccaca tcaacggcca cgacgacggc
     3481 gacatacgcc aatggcaatc ccaattctaa cgccaatccc agccagagcc agccgtccaa
     3541 tgcgctgttt tgcgagcagg tgaccacagt gacaaatctc ttcgagaagt ggaacgactg
     3601 cgagcgaacc gtcgtgatgt acgcgctgct caagcggctg cgctacccga gcctcaagtt
     3661 tctacagtat tctatcgaca gcaacctgac ccagaacctg ggcacctcgc agaccaatct
     3721 gagcagcgtg gttattgaca tcaatgccaa caatccggtg tacctgcaga atctactcaa
     3781 cgcctacaag acgttccagc catgtgacct gctcgacgcc atgtcctcct cgtcgtccga
     3841 caaggactcg atgccatgct acggcagtga cttccagatc accacatcgg cgcagtgtga
     3901 cgagcggaag ctgtatgccc gcaaggagga cattctgcat gaagtactga acatgctgcc
     3961 tctgctgaag ccgggcaatg aggaggccaa gcttatctac ctgaccctta taccagttgc
     4021 cgtcaaggac accatgcagc aaattgtgcc cacggagttg gtgcagcaga tcttctcgta
     4081 cctactcatc catccagcta tcaccagcga ggacagacgt tcgctcaaca tttggctgcg
     4141 tcacttggag gatcatatcc aagcggctgc ggcgggcctg acaaatcgca gttacttcct
     4201 gcagccctcg ccgcaactgg tcgctggtgg tagctcaaca ggcagtggta gctgttcctc
     4261 ttcggcgacc agctcttcga cagcatcctg ttcgtcggtg gcctcatcct cgttgtgccc
     4321 cgcaagcggc agccgttcct cacgcaccaa cgactggcaa acgatcgccc cgcccagcaa
     4381 gcaattgcaa aacaaattgg ccggtgattg gcgtggcaat ggaggcggca gctccagcgg
     4441 gtcgatcaat ccactctgtg ataatcttaa tggtattacc ctgaacgaat tagcttctag
     4501 tcagaacagt ctcggcctgt cgctggaggg cagctcttcc ctggtcaacg gtgtggtcgc
     4561 tggtgcaggc tctatgttgg gaatcgctgg tggcgacgat catgacacgt catttagcaa
     4621 gaatggcacg gagattctcg attttgaccc ggtgacggcg gacatgggtg aggcctgtag
     4681 tctggccagc agtagccttt gtggacgtaa cggtggaaat cctgttgagg atcgatcgca
     4741 acctccgccg aatcttcagc agcaattgct tcagcctcct ccctatgcct cgatccttat
     4801 gggaaacgtg ggcgatcagt ttggagagat caaccggtgg agcctggaca gcaagattgc
     4861 agccctgaag acgcggcgct ctaacagcct caccacgcaa acgatctcta gctgttccag
     4921 ctcgtctaac tcatcggtga tcacggtcaa tgacaactgc tccaattcaa cggagaatct
     4981 ggcacagttt gccaacaagc cccgtagttt ttccttatcg attgagcatc agcgcggcgc
     5041 cttgatgaac agcggttccg atactcgtct ggatgaacgc tcgagtctag agggcccgcg
     5101 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     5161 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     5221 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     5281 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     5341 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     5401 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     5461 ctacaaatgt ggtatggctg attatgatca gtcgaccgat gcccttgaga gccttcaacc
     5521 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     5581 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     5641 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     5701 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     5761 ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc
     5821 gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg
     5881 cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat
     5941 cgctcgcggc tcttaccagc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6001 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6061 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6121 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6181 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6241 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6301 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6361 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6421 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6481 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6541 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6601 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6661 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6721 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     7081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7381 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7741 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7801 ggcgtatcac gaggcccttt cgt
//
