LOCUS       sequence-102197-        5247 bp    DNA     circular SYN 15-APR-2025
DEFINITION  docking site transgenesis plasmid for GoldenGate Cloning and insect 
            transgenesis, DsRed-NLS marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5247)
  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 5247)
  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"; journalName: "G3 
            (Bethesda). 2015 Apr 13. pii: g3.115.016808. doi: 
            10.1534/g3.115.016808."
COMMENT     Sequence Label: pDSARN
FEATURES             Location/Qualifiers
     source          1..5247
                     /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             2232..2906
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2412..2432)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2824..2847
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2916..2936
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3077..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3259..3292
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3359..3458)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3489..3507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3512..3528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3630..3649)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3641..4450
                     /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(3708..3727)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4597..5185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5086..5105
                     /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 catggcctcc tccgagaacg tcatcaccga gttcatgcgc ttcaaggtgc
     2281 gcatggaggg caccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2341 cctacgaggg ccacaacacc gtgaagctga aggtgaccaa gggcggcccc ctgcccttcg
     2401 cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg
     2461 ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2521 tgaacttcga ggacggcggc gtggcgaccg tgacccagga ctcctccctg caggacggct
     2581 gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtgatgc
     2641 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     2701 tgaagggcga gacccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     2761 tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggacg
     2821 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgca
     2881 ccgagggccg ccaccacctg ttcctgagat ctcgacccaa gaaaaagcgg aaggtggagg
     2941 acccgtaaga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     3001 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3061 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3121 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3181 actcatcaat gtatcttatc atgtctggat cccgtttgac ggtatcgata agcttgatgg
     3241 ggatccggaa cccttaatta ccgttcgtat aatgtatgct atacgaagtt attaggtccc
     3301 tcgacctgca gcccgtacgg cgcgcctaga gcggccgcca ccgcggtgga gctcgagtac
     3361 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     3421 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg
     3481 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3541 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     3601 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3661 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3721 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3781 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3841 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3901 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3961 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4021 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4081 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4141 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4201 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4261 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4321 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4381 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     4441 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     4501 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     4561 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4621 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4681 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4741 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4801 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4861 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4921 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4981 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5041 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5101 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5161 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5221 cttttgctgg ccttttgctc acatgtt
//
