LOCUS       sequence-288359-        8149 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Insect Expression of DmBelle_1-694.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8149)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8149)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-DmBelle_1-694_B
FEATURES             Location/Qualifiers
     source          1..8149
                     /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             5434..5475
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5485..5502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5525..5542)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5575..5709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5599..5618)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5653..5672
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6331..6919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6411..6430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7090..7881,7882..7950))
                     /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     7713..7732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7951..8055)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8123..8141
                     /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 ttgatgagta atgctattaa ccaaaatggc acaggtctag agcagcaagt
     3361 tgctggtctg gacttgaatg gcggctctgc tgactacagc ggccccataa ccagcaagac
     3421 ttccactaac tcggtcaccg gtggtgtgta tgtgcccccg caccttcgtg gtggtggtgg
     3481 caataacaac gcagcggacg cagagagcca gggccaagga cagggccagg gtcagggctt
     3541 cgactccaga tccgggaatc cgcgccagga gactagggac cctcaacagt cgcgcggagg
     3601 cggaggcgaa taccgcagag gcggcggcgg tggtggtcgc ggcttcaatc gccagagcgg
     3661 cgactacggt tacggaagcg gcggcggagg acgacgtgga ggcggtggac gtttcgagga
     3721 caactacaac ggtggtgaat tcgattcgcg tcgcggcggt gactggaatc gcagcggcgg
     3781 aggcggcgga ggaggtcgag gatttggacg cggaccatcg taccgcggcg gtggcggcgg
     3841 aagcggtagc aacctcaacg agcagactgc cgaggatggc caggcacagc agcagcagca
     3901 gccgcgcaac gatcgttggc aggagccaga acgtcctgcc ggattcgatg gcagcgaggg
     3961 cggacagtct gccggcggca acagaagcta caacaatcgc ggcgagcgcg gaggcggcgg
     4021 ttacaacagc cgctggaagg agggcggcgg ctcaaacgtc gactacacaa agctgggcgc
     4081 ccgagatgag cgcctggagg tggagctgtt cggcgtgggc aatacgggaa tcaattttga
     4141 caaatacgag gatatacccg tggaggcgac gggccagaat gtgcccccga acatcacatc
     4201 gttcgatgat gtgcagctga cggagattat ccgcaacaac gtggccttag cacgttatga
     4261 caaaccgaca ccggtgcaga agcacgccat tccgatcatt atcaatggcc gggatctaat
     4321 ggcctgcgcc cagactggat ccggcaagac ggccgccttc ctggtgccga ttctcaacca
     4381 gatgtacgag ctgggacatg tgcccccgcc gcaaagcacc cggcagtaca gccggcgcaa
     4441 gcagtatccg ttgggtctgg tgctggcgcc gacccgcgaa ctggccaccc agatctttga
     4501 ggaggccaag aagttcgcct atcgctcccg gatgcgcccg gcagtgctgt acggcggaaa
     4561 caatactagc gagcagatgc gcgagctgga ccgcggttgc catctgattg tggccacacc
     4621 cggtcgccta gaggacatga tcacacgcgg aaaggtggga ctggagaaca ttcgattcct
     4681 tgtactggat gaggctgatc gtatgttgga catgggtttc gagccgcaga tccgtcgcat
     4741 cgtcgaacag ctaaacatgc cgccaaccgg acagcgccag acgcttatgt tctcggccac
     4801 attccccaag cagatccagg agttagccag cgatttcctc agcaactaca ttttcttggc
     4861 cgttggtcgt gtgggctcca catctgagaa cattacgcag acaatcctgt gggtctatga
     4921 acccgacaag cgctcgtatc tgctggatct gctctcgtct atccgcgatg gccccgaata
     4981 caccaaagac agcctaacac tgatctttgt ggagactaag aagggtgcag actcgctgga
     5041 ggagttcctg taccagtgca atcatcccgt cactagcatc cacggtgatc gcacgcagaa
     5101 ggagcgcgag gaggcactgc gctgcttccg atcgggcgac tgccccatcc tggtggccac
     5161 agccgtggcc gctcgtggcc tggacattcc gcacgttaag cacgttatta actttgacct
     5221 gccctcggat gtggaggagt atgtccaccg tatcgggcgt accggacgta tgggtaatct
     5281 tggtgtggcc acctcctttt tcaacgaaaa gaaccgcaac atttgctccg atctcttgga
     5341 gctgctgatc gaaacgaagc aggagatccc cagtttcatg gaggatatga gttcgtaggc
     5401 ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc
     5461 ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga
     5521 tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca
     5581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     5641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     5701 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     5761 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg
     5821 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     5881 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     5941 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     6001 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     6061 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
     6121 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
     6181 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
     6241 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagccagc aaaaggccag
     6301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7381 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     7441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7681 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     7741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8101 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgt
//
