LOCUS       62056_3460        9937 bp DNA     circular SYN 27-JAN-2021
DEFINITION  Cloning vector pBAC_attBs_fle-3miR, complete sequence.
ACCESSION   MW147152
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9937)
  AUTHORS   Krzywinska E, Ferretti L, Li J, Li JC, Chen CH, Krzywinski J.
  TITLE     Femaleless Controls Sex Determination and Dosage Compensation 
            Pathways in Females of Anopheles Mosquitoes
  JOURNAL   Curr Biol (2021) In press
  PUBMED    33417880
REFERENCE   2  (bases 1 to 9937)
  AUTHORS   Krzywinski J.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2020) Vector Molecular Biology Group, The 
            Pirbright Institute, Ash Road, Pirbright GU24 0NF, United Kingdom
REFERENCE   3  (bases 1 to 9937)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Biol 
            (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-OCT-2020) Vector Molecular Biology Group, The Pirbright 
            Institute, Ash Road, Pirbright GU24 0NF, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..9937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    122..242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(274..307)
                     /label=attB
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et
                     al., 2000)"
     misc_feature    313..1139
                     /label=piggyBac right arm
                     /note="piggyBac right arm"
     protein_bind    complement(1148..1181)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             1531..2187
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     rep_origin      2314..2902
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3658..3691
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    3702..4402
                     /label=piggyBac left arm
                     /note="piggyBac left arm"
     protein_bind    4408..4441
                     /label=attB
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et
                     al., 2000)"
     gene            4472..6434
                     /gene="AGAP001971"
                     /label=AGAP001971
     regulatory      4472..6434
                     /gene="AGAP001971"
                     /label=Anopheles gambiae polyubiquitin
                     /note="Anopheles gambiae polyubiquitin"
                     /regulatory_class="promoter"
     misc_RNA        6445..7092
                     /label=cluster of 3 microRNAs targeting the fle gene
                     /note="cluster of 3 microRNAs targeting the fle gene"
     regulatory      7106..7335
                     /label=SV40
                     /note="SV40"
                     /regulatory_class="terminator"
     gene            7349..7946
                     /gene="AGAP004395"
                     /label=AGAP004395
     regulatory      7349..7946
                     /gene="AGAP004395"
                     /regulatory_class="promoter"
     CDS             7953..8549
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     regulatory      8559..8788
                     /label=SV40
                     /note="SV40"
                     /regulatory_class="terminator"
     gene            8789..9217
                     /gene="AGAP003430"
                     /label=AGAP003430
     regulatory      8789..9217
                     /gene="AGAP003430"
                     /regulatory_class="promoter"
     CDS             9218..9934
                     /label=EGFP
                     /note="enhanced GFP"
ORIGIN
        1 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
       61 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
      121 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
      181 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
      241 tttccggagc gatcgcatgg gtgaggtgga gtacgcgccc ggggagccca agggcacgcc
      301 ctggcacccg cacctgcagt aggaagacga ataggtggcc tatggcatta ttgtacggaa
      361 tgataaacat tgcctgcata aattctttta ttatatacag ccataatgtc agtagcaagg
      421 gagaaaaggt tcaaagtcgc aaaaaattta tgagaaacct ttacatgagc ctgacgtcat
      481 cgtttatgcg taagcgttta gaagctccta ctttgaagag atatttgcgc gataatatct
      541 ctaatatttt gccaaatgaa gtgcctggta catcagatga cagtactgaa gagccagtaa
      601 tgaaaaaacg tacttactgt acttactgcc cctctaaaat aaggcgaaag gcaaatgcat
      661 cgtgcaaaaa atgcaaaaaa gttatttgtc gagagcataa tattgatatg tgccaaagtt
      721 gtttctgact gactaataag tataatttgt ttctattatg tataagttaa gctaattact
      781 tattttataa tacaacatga ctgtttttaa agtacaaaat aagtttattt ttgtaaaaga
      841 gagaatgttt aaaagttttg ttactttata gaagaaattt tgagtttttg ttttttttta
      901 ataaataaat aaacataaat aaattgtttg ttgaatttat tattagtatg taagtgtaaa
      961 tataataaaa cttaatatct attcaaatta ataaataaac ctcgatatac agaccgataa
     1021 aacacatgcg tcaattttac gcatgattat ctttaacgta cgtcacaata tgattatctt
     1081 tctagggtta aataatagtt tctaattttt ttattattca gcctgctgtc gtgaataccc
     1141 cacttcagaa gttcctatac tttctagaga ataggaactt cactatagag tcgaataagg
     1201 gcgacacccc ctaattagcc cgggcgaaag gcccagtctt tcgactgagc ctttcgtttt
     1261 atttgatgcc tggcagttcc ctactctcgc atggggagtc cccacactac catcggcgct
     1321 acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc tactgccgcc
     1381 aggcaaaggg gagagacgcg catcagaatt ggttaattgg ttgtaacact gacccctatt
     1441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1501 atgcttcaat aatattgaaa aaggaagaat atggagaaaa aaatcactgg atataccacc
     1561 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     1621 tgtacctata accagaccgt tcagctggat attacggcct ttttaaaaac cgtaaagaaa
     1681 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     1741 ccagaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccca
     1801 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     1861 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     1921 ctggcctatt tccctaaagg atttattgag aatatgtttt tcgtgtcagc caatccctgg
     1981 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     2041 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     2101 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     2161 tactgcgatg agtggcaggg cggggcgtaa cgtctcgaaa atacgctgac ttgacgggac
     2221 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     2281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2461 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     2521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2941 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3001 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3061 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     3121 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     3181 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     3241 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     3301 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     3361 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     3421 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     3481 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     3541 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     3601 cagtatttat tatgcattta gaataaattt tgtgtcgccc ttattcgact cactatagaa
     3661 gttcctattc tctagaaagt ataggaactt cacttcattt tcgcaaatct tttttaaccc
     3721 tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct
     3781 aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg
     3841 tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag
     3901 tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta
     3961 tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata
     4021 gatatcgtga ctaatatata ataaaatggg tagttcttta gacgatgagc atatcctctc
     4081 tgctcttctg caaagcgatg acgagcttgt tggtgaggat tctgacagtg aaatatcaga
     4141 tcacgtaagt gaagatgacg tccagagcga tacagaagaa gcgtttatag atgaggtaca
     4201 tgaagtgcag ccaacgtcaa gcggtagtga aatattagac gaacaaaatg ttattgaaca
     4261 accaggttct tcattggctt ctaacagaat cttgaccttg ccacagagga ctattagagg
     4321 taagaataaa cattgttggt caacttcaaa gtccacgagg cgtagccgag tctctgcact
     4381 gaacattgtc agatccgaga tctgcgggtg ccagggcgtg cccttgggct ccccgggcgc
     4441 gtactccacc tcacccatgg gacccacgcg tcagtccact tatggaatca tcattgtaat
     4501 atgagaagaa aagaaaaaaa acttttgaaa cttcgcaaag gatttcgatc gtgaacttca
     4561 aatgtctcac tcaagtgagc taaaatttag ctcactggtt gaattttgct gacaagtgat
     4621 cacttattga tgcatataat gtataataaa atgtgttcaa ttcgtaaaag cacaaacaag
     4681 aacagtaaat cattgcgatt tgtctgactg aattttatga aatgaatttt gttataataa
     4741 ttatttcacg tacaaaaatc cgcttctcat ttctctcgct gcaaactcac tcgcaaatca
     4801 gcacgtgtaa actcaacttt agcgcagaac atttgacgca cacaaatgat actgttgttg
     4861 tatttgttgc cgacgaaaag gctataaaaa cgaggaattc gcggaagcag accgagccaa
     4921 gttaaagcaa aaccagacaa gcagtgctac agtacggacg cgcaggcagg acgtgtgaag
     4981 cagaatatca attgtgcttt gtggagaaaa gaaaattatt attattaact gtgttgtgta
     5041 aaaggtaatg caactactgt atgcttgtgt tgataagttc tgcattaaag ttttctgaca
     5101 agcgcgtgcg aaaggcagag attggtttta attgtttggg gtaactaggt ttttcccaag
     5161 atggcgtaat gacgcagtga gctgtgactg tggtgcaagc aatttatcat ttgctcaatc
     5221 gctcaacgta tttaattagt gcgtgttttc gcgaaatagt tgtgtatttc gtaagaaaat
     5281 ctttatattt catcgcaaat cttttgtgtt cctatattag tgttagtata gagaaggttc
     5341 gtagttgcaa aatgtgttca aattcaatca ttgtgattgt ataaaacatc aaatttaaat
     5401 ctttattaag tttacaagta aagtatgagg attttaaatg tactatagtg taggttattt
     5461 tagtgaaagt gagttagaaa gctaattttt tttttcgtgt acggtttcta tgaaccagtt
     5521 atgcgcagct tgcgttgtat cccttctgcg aacaaagaga tgattatcgt aaatgaaaaa
     5581 tggcgtcatc taatgccatg aactcgatgt acagtgtaca tttcagttag tttttgttat
     5641 ctgttgaaat agaaaacaaa acgaattata aaagactaac cagaagtgtt tgtttcaatt
     5701 tacttaatgg attctttagt tacaaaaaat ataaaaaaac tttattataa tataataaaa
     5761 atcttttatc ttttaaaata agactagtta aaaaattcac atcaaattta tattcctgta
     5821 tgatttaatg gtgaaacaaa ttatcttagt tattaaggcc tcgaacacgc tcgaacatgt
     5881 tcaagtgctt gcgtgtggtt gccattgaat gactcatttt cacttcgtag attgcaagtg
     5941 tctttaggct aaactaaact tacaagctga aaagcaaacc gctgttttcg attaattaat
     6001 taaattaaat gctctgaatg ttcttgttaa acacattttt gatgttacgc acactccttt
     6061 cgtcgtgaat ttatttatag attaggtcac tgtgaggggt tacaatctag aagtttcgat
     6121 tggaaaagta aattgataca agtttttcta gactgttcct tccctcccac actcccaccc
     6181 cttccaccga attcagaata tattcagtgg tgatgcaata tacctcaaac atcacttggg
     6241 atttatgatg aaatagttac gttgtttaac gtaggtttaa ttggattaag atattataga
     6301 aggagcacaa acgagagtgc ataacatagc attcttgcag taatgtagtg cctaagacgt
     6361 tgcttgctat gatgtgatga actgagcatt gcacaatgct ataatgttgt tttcgtttaa
     6421 atttacagat aaaatatgag atcttttaaa gtccacaact catcaaggaa aatgaaagtc
     6481 aaagttggca gcttacttaa acttaatcac agcctttaat gtaaagacat caagctaatc
     6541 tacgtaagtt aatataccat atctacgtag attagcttga tgtctttgta cctaaagtgc
     6601 ctaacatcat tatttaattt tttttttttt tggcacacga ataaccatgc cgttttggat
     6661 cttttaaagt ccacaactca tcaaggaaaa tgaaagtcaa agttggcagc ttacttaaac
     6721 ttaatcacag cctttaatgt gcatcgtaca gacatcaaca cgtaagttaa tataccatat
     6781 ctacgtgttg atgtctgtac gatgcgtacc taaagtgcct aacatcatta tttaattttt
     6841 tttttttttg gcacacgaat aaccatgccg ttttggatct tttaaagtcc acaactcatc
     6901 aaggaaaatg aaagtcaaag ttggcagctt acttaaactt aatcacagcc tttaatgtgg
     6961 atacgctacc aagactcaat taagttaata taccatatct aattgagtct tggtagcgta
     7021 tccgtaccta aagtgcctaa catcattatt taattttttt tttttttggc acacgaataa
     7081 ccatgccgtt ttggatccgg ccggcctaga tcataatcag ccataccaca tttgtagagg
     7141 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     7201 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7261 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7321 tcatcaatgt atcttggcct gacaggccgt acgatgccaa ctttgctggc tatattcagt
     7381 cttgggttga gcgcttcccc gatacgaaaa tcgacgacat tctggaacgc atccatgcga
     7441 cggataagca atattttgaa ggattgtatt aaacgtctgc gtccaaatgc gatatgcaaa
     7501 taagattcaa gctgaataaa gaaggttttt gaacaagtat ttagtttccg cttgctgtgc
     7561 gtttatttca ctgcttcacg cagcacaaca gcatgattaa tatgtatgta aatagaagca
     7621 gactcttgtg gaggtagtga acttttccat agcacaacgt ctttgtagat ctaaaacagt
     7681 gatctattat ttcgcaagca gcgtacaccg aacacggtat tgaaatttca aattgacatg
     7741 ttgacagttg ccattattgc agaccgaaga gttccgaatg ttgccgaagg ttgacagcgc
     7801 ggctcaccag caccgtacaa agcttgggca aaacggatgt gtttttttca ttctcttgca
     7861 agtttccaaa cgtggtgaga gtcgcgcatc catttgcttt gctgaagaaa atccggaaaa
     7921 agccaactaa ccgcacacaa atatggccta ggatgaccga gtacaagccc acggtgcgcc
     7981 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
     8041 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
     8101 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     8161 gcgccgcggt ggcggtctgg accacgccgg aaagcgtcaa agcgggggcg gtgttcgccg
     8221 agatcggccc gcgcatggcc aagttgagcg gttcccggct ggccgcgcag caacaaatgg
     8281 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     8341 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     8401 cggccgagcg cgccggggtg cccgccttcc tggaaacttc cgcgccccgc aacctcccct
     8461 tctacaagcg gctcggcttc accgtcaccg ccaacgtcaa ggtgcccgaa ggaccgcgca
     8521 cctggtgcat gacccgcaag cccggtgcct gagctagcct agatcataat cagccatacc
     8581 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     8641 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     8701 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     8761 ggtttgtcca aactcatcaa tgtatcttgt tgtgcataat cagcgtcgtg tatgcaaaac
     8821 gaagcagcag tagtagagta tcagctcgac aaatgtttca tacatcttta tatacttgga
     8881 atgttcggta gaatatttat tttaataaat ataagtacat gtaaaagctg ttgcttcttt
     8941 ttttgttctt tacgaatcaa ttaaaaataa ttaaaagtca tactgaaatt ttaaacagaa
     9001 atcgagcacg aatactcgtt tgtcgtagct ttttcaccgg cagattcgag ctgctgaaaa
     9061 tgaaggcaga atcgagccca ccgcacagcc ctgcgcaaag tgtcacgtga ggaacgggac
     9121 ggaaagtgga acaaaaatca attttctagc aaccacaata acgtcatatg atcacaagtg
     9181 caaattcgga gggcagataa cttcactgga cgacaagatg gtgagcaagg gcgaggagct
     9241 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     9301 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     9361 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     9421 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     9481 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     9541 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     9601 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     9661 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     9721 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     9781 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     9841 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     9901 cgggatcact ctcggcatgg acgagctgta caagtaa
//
