LOCUS       sequence-112603-        5143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  docking site transgenesis plasmid for GoldenGate Cloning and insect 
            transgenesis, GFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  AUTHORS   Volohonsky G, Terenzi O, Soichot J, Naujoks DA, Nolan T, Windbichler
            N, Kapps D, Smidler AL, Vittu A, Costa G, Steinert S, Levashina EA, 
            Blandin SA, Marois E
  TITLE     Tools for Anopheles gambiae Transgenesis.
  JOURNAL   G3 (Bethesda). 2015 Apr 13. pii: g3.115.016808. doi: 
            10.1534/g3.115.016808.
  PUBMED    25869647
REFERENCE   2  (bases 1 to 5143)
  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"; journalName: "G3 
            (Bethesda). 2015 Apr 13. pii: g3.115.016808. doi: 
            10.1534/g3.115.016808."
COMMENT     Sequence Label: pDSAG
FEATURES             Location/Qualifiers
     source          1..5143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     271..287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(337..461)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    490..511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(526..531,532..549,550..556)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    564..580
                     /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)."
     primer_bind     569..591
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     588..604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             600..1175
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     623..643
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        625..643
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    656..763
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     680..696
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     680..696
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(730..746)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(731..747)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(772..790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(800..816)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      958..1338
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1045..1064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1255..1276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1502..1584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1642..1711
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     regulatory      2226..2235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2232..2234,2235..2237,2238..2951)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2232..2234,2235..2237,2238..2948)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2277..2298)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2538..2557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2885..2906
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3073..3194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3255..3354)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3385..3403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3408..3424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3526..3545)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3537..4346
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3604..3623)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4493..5081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4982..5001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      301 ccctacaggt cactaatacc atctaagtag ttgattcata gtgactggat atgttgtgtt
      361 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
      421 tcattttacg tttctcgttc aacttttcta tacaaagttg gtaccggatc cagagacccg
      481 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
      541 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
      601 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
      661 ccaccgcggt ggcggccgct ctagaactag tggatccccc gggctgcagg aattcgatat
      721 caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg ccctatagtg
      781 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
      841 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
      901 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
      961 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1021 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1081 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1141 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1201 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1261 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1321 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1381 cgggtctccg cttctagaca taatcagcca taccacattt gtagaggttt tacttgcttt
     1441 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1501 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1561 aaataaagca tttttcttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1621 ctcgacgatg taggtcacag tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg
     1681 ctccccgggc gcgtactcca cctcacccat ctggtccatc atgatgaacg ggtcgaggtg
     1741 gcggtagttg atcccggcga acgcgcggcg caccgggaag ccctcgccct cgaaaccgct
     1801 gggcgcggtg gtcacggtga gcacgggacg tgcgacggcg tcggcgggtg cggatacgcg
     1861 gggcagcgtc agcgggttct cgacggtcac ggcgggcaat tcctgcagac ttccggtatc
     1921 tcgcgtttgt ttgatcgcac ggttcccaca atggttaatt cgagctcgcc cggggatcta
     1981 attcaattag agactaattc aattagagct aattcaatta ggatccaagc ttatcgattt
     2041 cgaaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat
     2101 tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct
     2161 gaacaagcta aacaatcggg gtaccgctag agtcgacggt accgcgggcc cgggatccac
     2221 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2281 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2341 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2401 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2461 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2521 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2581 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2641 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2701 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2761 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2821 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2881 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2941 tgtacaagta aagcggccgc gactctagat caaatcagcc ataccacatt tgtagaggtt
     3001 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3061 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3121 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3181 atcaatgtat cttaaagctt atcgatacgc gtacggcgcg cctagagcgg ccgccaccgc
     3241 ggtggagctc gagtacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3301 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     3361 agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     3421 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     3481 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     3541 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     3601 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     3661 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     3721 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     3781 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     3841 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     3901 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     3961 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     4021 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     4081 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     4141 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     4201 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     4261 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     4321 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     4381 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     4441 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4501 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4561 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4621 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4681 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4741 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4801 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4861 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4921 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4981 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5041 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5101 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
