LOCUS       sequence-112606-        4884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  docking site transgenesis plasmid for Gateway Cloning and insect 
            transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4884)
  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 4884)
  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: pattB-RfB2
FEATURES             Location/Qualifiers
     source          1..4884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    665..789
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(830..1135)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(932..951)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1477..2136)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2051..2070
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2190..2196,2197..2214,2215..2220))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2245..2369)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    2406..2475
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(2694..2712)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2733..2749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2733..2749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2746..2768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2757..2773
                     /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(2781..2787,2788..2805,2806..2811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2826..2847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2964..2981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3135..3723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3215..3234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4192..4572)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     4519..4538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4757..4861)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 atcaaccact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa atatcaatat
      721 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
      781 gtcactatgg tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc
      841 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
      901 cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
      961 catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg
     1021 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
     1081 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
     1141 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
     1201 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
     1261 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
     1321 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
     1381 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
     1441 ctgttatctg gcttttagta agccggatcc agatctttac gccccgccct gccactcatc
     1501 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     1561 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     1621 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     1681 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     1741 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     1801 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     1861 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     1921 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     1981 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     2041 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     2101 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     2161 tctcgacgga tcctaactca aaatccacac attatacgag ccggaagcat aaagtgtaaa
     2221 gcctggggtg cctaatgcgg ccgccatagt gactggatat gttgtgtttt acagtattat
     2281 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     2341 tctcgttcag cttttttgta caaacttgtt gatgatccac tagtgtcgac gatgtaggtc
     2401 acggtctcga agccgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
     2461 tccacctcac ccatctggtc catcatgatg aacgggtcga ggtggcggta gttgatcccg
     2521 gcgaacgcgc ggcgcaccgg gaagccctcg ccctcgaaac cgctgggcgc ggtggtcacg
     2581 gtgagcacgg gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag cgtcagcggg
     2641 ttctcgacgg tcacggcggg catgtcgaca ctagttctag ccagcttttg ttccctttag
     2701 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2821 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2881 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2941 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3001 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3061 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3121 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3181 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3241 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3301 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3361 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3421 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3481 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3541 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3601 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3661 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3721 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3781 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3841 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3901 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3961 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4021 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4081 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4141 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4201 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4261 ggttcccaac gatcnaaggc gagttacatg atncccccat gttgtgcaaa aaagcggtta
     4321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4861 gcacatttcc ccgaaaagtg ccac
//
