LOCUS       sequence-180860-        6865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EYFP in eyes and contains an attP landing site for 
            site-specific attB integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6865)
  AUTHORS   Stern DL, Crocker J, Ding Y, Frankel N, Kappes G, Kim E, Kuzmickas 
            R, Lemire A, Mast JD, Picard S
  TITLE     Genetic and Transgenic Reagents for Drosophila simulans, D. 
            mauritiana, D. yakuba, D. santomea, and D. virilis.
  JOURNAL   G3 (Bethesda). 2017 Apr 3;7(4):1339-1347. doi: 
            10.1534/g3.116.038885.
  PUBMED    28280212
REFERENCE   2  (bases 1 to 6865)
  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"; doi: "10.1534/g3.116"; 
            journalName: "G3 (Bethesda)"; date: "2017-04-3- 3"; volume: "7"; 
            issue: "4"; pages: "1339-1347"
COMMENT     Sequence Label: pBac{3XP3::EYFP,attP}
FEATURES             Location/Qualifiers
     source          1..6865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..720)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    843..964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(880..899)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     934..953
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1159..1258
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     repeat_region   2017..2051
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2232..2248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2232..2248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2245..2267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2256..2272
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2280..2286,2287..2304,2305..2310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2325..2346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2463..2480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2634..3222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2714..2733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3393..4184,4185..4253))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4016..4035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4254..4358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4426..4444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4482..4504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4604..4623
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4817..4839
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4831..4848
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4832..4848
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   5552..5614
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      join(6860..6865,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      721 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
      781 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
      841 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      901 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      961 cttaaagctt atcgatacgc gtacggcgcg cctaggccgg ccgaattcga atggccatgg
     1021 gacgtcgacc tgaggtaatt ataacccggg ccctatatat ggatccaatt gcaatgatca
     1081 tcatgacaga tctgtactga cggacacacc gaagccccgg cggcaaccct cagcggatgc
     1141 cccggggctt cacgttttcc caggtcagaa gcggttttcg ggagtagtgc cccaactggg
     1201 gtaacctttg agttctctca gttgggggcg tagggtcgcc gacatgacac aaggggttgt
     1261 gaccggggtg gacacgtacg cgggtgctta cgaccgtcag tcgcgcgagc gcgatctaga
     1321 gtcgtcgacg tcaaaatcac cacttttgaa gcgttgaaac caccgttcac acagatctga
     1381 caatgttcag tgcagagact cggctacgcc tcgtggactt tgaagttgac caacaatgtt
     1441 tattcttacc tctaatagtc ctctgtggca aggtcaagat tctgttagaa gccaatgaag
     1501 aacctggttg ttcaataaca ttttgttcgt ctaatatttc actaccgctt gacgttggct
     1561 gcacttcatg tacctcatct ataaacgctt cttctgtatc gctctggacg tcatcttcac
     1621 ttacgtgatc tgatatttca ctgtcagaat cctcaccaac aagctcgtca tcgctttgca
     1681 gaagagcaga gaggatatgc tcatcgtcta aagaactacc cattttatta tatattagtc
     1741 acgatatcta taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa
     1801 taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt
     1861 cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc
     1921 acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg
     1981 cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta
     2041 tctttctagg gttaaaaaag atttgcgctt tactcgacct aaactttaaa cacgtcatag
     2101 aatcttcgtt tgacaaaaac cacattgtgg ccaagctgtg tgacgcgacg cgcgctaaag
     2161 aatggcaaac caagtcgcgc gagcgtcgac tctagaggat ccccgggtac cgagctcgaa
     2221 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2281 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2341 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3061 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4381 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4441 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4501 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4561 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4621 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4681 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     4741 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     4801 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctttgt
     4861 ttaaaaatat aacaaaattg tgatcccaca aaatgaagtg gggcaaaatc aaataattaa
     4921 tagtgtccgt aaacttgttg gtcttcaact ttttgaggaa cacgttggac ggcaaatccg
     4981 tgactataac acaagttgat ttaataattt tagccaacac gtcgggctgc gtgttttttg
     5041 ccgacgcgtc tgtgtacacg ttgattaact ggtcgattaa actgttgaaa taatttaatt
     5101 tttggttctt ctttaaatct gtgatgaaat tttttaaaat aactttaaat tcttcattgg
     5161 taaaaaatgc cacgttttgc aacttgtgag ggtctaatat gaggtcaaac tcagtaggag
     5221 ttttatccaa aaaagaaaac atgattacgt ctgtacacga acgcgtatta acgcagagtg
     5281 caaagtataa gagggttaaa aaatatattt tacgcaccat atacgcatcg ggttgatatc
     5341 gttaatatgg atcaatttga acagttgatt aacgtgtctc tgctcaagtc tttgatcaaa
     5401 acgcaaatcg acgaaaatgt gtcggacaat atcaagtcga tgagcgaaaa actaaaaagg
     5461 ctagaatacg acaatctcac agacagcgtt gagatatacg gtattcacga cagcaggctg
     5521 aataataaaa aaattagaaa ctattattta accctagaaa gataatcata ttgtgacgta
     5581 cgttaaagat aatcatgcgt aaaattgacg catgtgtttt atcggtctgt atatcgaggt
     5641 ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca tactaataat
     5701 aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaaaca aaaactcaaa
     5761 atttcttcta taaagtaaca aaacttttaa acattctctc ttttacaaaa ataaacttat
     5821 tttgtacttt aaaaacagtc atgttgtatt ataaaataag taattagctt aacttataca
     5881 taatagaaac aaattatact tattagtcag tcagaaacaa ctttggcaca tatcaatatt
     5941 atgctctcga caaataactt ttttgcattt tttgcacgat gcatttgcct ttcgccttat
     6001 tttagagggg cagtaagtac agtaagtacg ttttttcatt actggctctt cagtactgtc
     6061 atctgatgta ccaggcactt catttggcaa aatattagag atattatcgc gcaaatatct
     6121 cttcaaagta ggagcttcta aacgcttacg cataaacgat gacgtcaggc tcatgtaaag
     6181 gtttctcata aattttttgc gactttgaac cttttctccc ttgctactga cattatggct
     6241 gtatataata aaagaattta tgcaggcaat gtttatcatt ccgtacaata atgccatagg
     6301 ccacctattc gtcttcctac tgcaggtcat cacagaacac atttggtcta gcgtgtccac
     6361 tccgccttta gtttgattat aatacataac catttgcggt ttaccggtac tttcgttgat
     6421 agaagcatcc tcatcacaag atgataataa gtataccatc ttagctggct tcggtttata
     6481 tgagacgaga gtaaggggtc cgtcaaaaca aaacatcgat gttcccactg gcctggagcg
     6541 actgtttttc agtacttccg gtatctcgcg tttgtttgat cgcacggttc ccacaatggt
     6601 taattcgagc tcgcccgggg atctaattca attagagact aattcaatta gagctaattc
     6661 aattaggatc caagcttatc gatttcgaac cctcgaccgc cggagtataa atagaggcgc
     6721 ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa
     6781 gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tcggggtacc gctagagtcg
     6841 acggtacgat ccaccggtcg ccacc
//
