LOCUS       sequence-102196-        5187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  docking site transgenesis plasmid for GoldenGate Cloning and insect 
            transgenesis, YFP-NLS marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  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 5187)
  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: pDSAYN
FEATURES             Location/Qualifiers
     source          1..5187
                     /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..2948)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     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"
     CDS             2958..2978
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3113..3234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3299..3398)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3429..3447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3452..3468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3570..3589)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3581..4390
                     /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(3648..3667)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4537..5125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5026..5045
                     /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 accttcggct acggcctgca gtgcttcgcc 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 taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2881 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2941 tgtacaagag atctcgaccc aagaaaaagc ggaaggtgga ggacccgtaa agcggccgcg
     3001 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3061 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3121 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3181 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaagctt
     3241 atcgatacgc gtacgtacgg cgcgcctaga gcggccgcca ccgcggtgga gctcgagtac
     3301 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     3361 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg
     3421 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3481 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     3541 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3601 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3661 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3721 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3781 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3841 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3901 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3961 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4021 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4081 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4141 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4201 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4261 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4321 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     4381 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     4441 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     4501 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4561 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4621 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4681 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4741 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4801 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4861 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4921 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4981 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5041 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5101 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5161 cttttgctgg ccttttgctc acatgtt
//
