LOCUS       sequence-102195-        5456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  docking site transgenesis plasmid for GoldenGate Cloning and insect 
            transgenesis, Puromycin resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5456)
  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 5456)
  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: pDSAP
FEATURES             Location/Qualifiers
     source          1..5456
                     /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)"
     promoter        2114..2661
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect 
                     cell expression"
     promoter        2697..2744
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        2736..2754
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2764..3363
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2764..2783)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3260..3280
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3375..3509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3568..3667)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3698..3716)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3721..3737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3839..3858)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3850..4659
                     /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(3917..3936)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4806..5394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5295..5314
                     /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 cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagtggat cccccgggct
     2101 gcaggaattc gatcatgatg ataaacaatg tatggtgcta atgttgcttc aacaacaatt
     2161 ctgttgaact gtgttttcat gtttgccaac aagcaccttt atactcggtg gcctccccac
     2221 caccaacttt tttgcactgc aaaaaaacac gcttttgcac gcgggcccat acatagtaca
     2281 aactctacgt ttcgtagact attttacata aatagtctac accgttgtat acgctccaaa
     2341 tacactacca cacattgaac ctttttgcag tgcaaaaaag tacgtgtcgg cagtcacgta
     2401 ggccggcctt atcgggtcgc gtcctgtcac gtacgaatca cattatcgga ccggacgagt
     2461 gttgtcttat cgtgacagga cgccagcttc ctgtgttgct aaccgcagcc ggacgcaact
     2521 ccttatcgga acaggacgcg cctccatatc agccgcgcgt tatctcatgc gcgtgaccgg
     2581 acacgaggcg cccgtcccgc ttatcgcgcc tataaataca gcccgcaacg atctggtaaa
     2641 cacagttgaa cagcatctgt tcgaattaat tcgggatctg ccgggctgca gcacgtgttg
     2701 acaattaatc atcggcatag tatatcggca tagtataata cgactcacta taggagggcc
     2761 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     2821 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     2881 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     2941 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     3001 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     3061 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     3121 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     3181 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     3241 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     3301 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     3361 tgactcgacc tcgaaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3481 tcatcaatgt atcttatcat gtctggatca tggggggggc ccggtacggc gcgcctagag
     3541 cggccgccac cgcggtggag ctcgagtacc cagctttctt gtacaaagtt ggcattataa
     3601 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     3661 ttatttgcca tccagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg
     3721 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     3781 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     3841 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     3901 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     3961 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     4021 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     4081 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     4141 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     4201 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     4261 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     4321 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     4381 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     4441 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     4501 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     4561 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     4621 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     4681 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     4741 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
