LOCUS       sequence-232708-        7929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express gRNA targeting Tra under dU6-3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7929)
  AUTHORS   Kandul NP, Liu J, Sanchez C HM, Wu SL, Marshall JM, Akbari OS
  TITLE     Transforming insect population control with precision guided sterile
            males with demonstration in flies.
  JOURNAL   Nat Commun. 2019 Jan 8;10(1):84. doi: 10.1038/s41467-018-07964-7.
  PUBMED    30622266
REFERENCE   2  (bases 1 to 7929)
  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: "Nat 
            Commun."; date: "2019-01-8"; volume: "10(1):84. doi"; pages: " 
            10.1038/s41467-018-07964-7"
COMMENT     Sequence Label: gRNA[Tra].1026G
FEATURES             Location/Qualifiers
     source          1..7929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     582..604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     596..613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     597..613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     620..639
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        620..638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(672..690)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1142..1213,1615..1888,1963..2617,2679..2994,3198..3329,
                     3400..4014)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4789..4822
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4881..5301
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     misc_RNA        5322..5397
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    5453..5522
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(5741..5759)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5780..5796)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5780..5796)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5793..5815)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5804..5820
                     /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(5828..5834,5835..5852,5853..5858))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5873..5894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6011..6028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6182..6770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6262..6281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6941..7732,7733..7801))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7564..7583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7802..7906)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttmcrt tcgccattca ggctgcgcaa
      481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      601 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggta cgtaccgggc
      661 ccctagtatg tatgtaagtt aataaaaccc atttttgcgg aaagtagata aaaaaaacat
      721 tttttttttt tactgcactg gatatcattg aacttatctg atcagtttta aatttacttc
      781 gatccaaggg tatttgatgt accaggttct ttcgattacc tctcactcaa aatgacattc
      841 cactcaaagt cagcgctgtt tgcctccttc tctgtccaca gaaatatcgc cgtctctttc
      901 gccgctgcgt ccgctatctc tttcgccacc gtttgtagcg ttacgtagcg tcaatgtccg
      961 ccttcagttg cattttgtca gcggtttcgt gacgaagctc caagcggttt acgccatcaa
     1021 ttaaacacaa agtgctgtgc caaaactcct ctcgcttctt atttttgttt gttttttgag
     1081 tgattggggt ggtgattggt tttgggtggg taagcagggg aaagtgtgaa aaatcccggc
     1141 aatgggccaa gaggatcagg agctattaat tcgcggaggc agcaaacacc catctgccga
     1201 gcatctgaac aatgtgagta gtacatgtgc atacatctta agttcacttg atctatagga
     1261 actgcgattg caacatcaaa ttgtctgcgg cgtgagaact gcgacccaca aaaatcccaa
     1321 accgcaattg cacaaacaaa tagtgacacg aaacagatta ttctggtagc tgttctcgct
     1381 atataagaca atttttgaga tcatatcatg atcaagacat ctaaaggcat tcattttcga
     1441 ctatattctt ttttacaaaa aatataacaa ccagatattt taagctgatc ctagatgcac
     1501 aaaaaataaa taaaagtata aacctacttc gtaggatact tcggggtact ttttgttcgg
     1561 ggttagatga gcataacgct tgtagttgat atttgagatc ccctatcatt gcagggtgac
     1621 agcggagcgg cttcgcagag ctgcattaac cagggcttcg ggcaggccaa aaactacggc
     1681 acgctccggc cacccagtcc gccggaggac tccggttcag ggagcggcca actagccgag
     1741 aacctcacct atgcctggca caatatggac atctttgggg cggtcaatca gccgggctcc
     1801 ggatggcggc agctggtcaa ccggacacgc ggactattct gcaacgagcg acacataccg
     1861 gcgcccagga aacatttgct caagaacggt gagtttctat tcgcagtcgg ctgatctgtg
     1921 tgaaatctta ataaagggtc caattaccaa tttgaaactc agtttgcggc gtggcctatc
     1981 cgggcgaact tttggccgtg atgggcagtt ccggtgccgg aaagacgacc ctgctgaatg
     2041 cccttgcctt tcgatcgccg cagggcatcc aagtatcgcc atccgggatg cgactgctca
     2101 atggccaacc tgtggacgcc aaggagatgc aggccaggtg cgcctatgtc cagcaggatg
     2161 acctctttat cggctcccta acggccaggg aacacctgat tttccaagcc atggtgcgga
     2221 tgccacgaca tctgacctat cggcagcgag tggcccgcgt ggatcaggtg atccaggagc
     2281 tttcgctcag caaatgtcag cacacgatca tcggtgtgcc cggcagggtg aaaggtctgt
     2341 ccggcggaga aaggaagcgt ctggcattcg cctccgaggc tctaaccgat ccgccgcttc
     2401 tgatctgcga tgagcccacc tccggactgg actcctttac cgcccacagc gtcgtccagg
     2461 tgctgaagaa gctgtcgcag aagggcaaga ccgtcatcct gaccattcat cagccgtctt
     2521 ccgagctgtt tgagctcttt gacaagatcc ttctgatggc cgagggcagg gtagctttct
     2581 tgggcactcc cagcgaagcc gtcgacttct tttcctagtg agttcgatgt gtttattaag
     2641 ggtatctagt attacataac atctcaactc ctatccagcg tgggtgccca gtgtcctacc
     2701 aactacaatc cggcggactt ttacgtacag gtgttggccg ttgtgcccgg acgggagatc
     2761 gagtcccgtg atcggatcgc caagatatgc gacaattttg ccattagcaa agtagcccgg
     2821 gatatggagc agttgttggc caccaaaaat ctggagaagc cactggagca gccggagaat
     2881 gggtacacct acaaggccac ctggttcatg cagttccggg cggtcctgtg gcgatcctgg
     2941 ctgtcggtgc tcaaggaacc actcctcgta aaagtgcgac ttattcagac aacggtgagt
     3001 ggttccagtg gaaacaaatg atataacgct tacaattctt ggaaacaaat tcgctagatt
     3061 ttagatagaa ttgcctgatt ccacaccctt cttagttttt ttcaatgaga tgtatagttt
     3121 atagttttgc agaagataaa taaatttcat ttaactcgcg aatattaatg agatgcgagt
     3181 aacattttaa tttgcagatg gttgccatct tgattggcct catctttttg ggccaacaac
     3241 tcacgcaagt gggtgtgatg aatatcaacg gagccatctt cctcttcctg accaacatga
     3301 cctttcaaaa cgtctttgcc acgataaatg taagtcatgt ttagaataca tttgcatttc
     3361 aataatttac taactttcta atgaatcgat tcgatttagg tgttcacctc agagctgcca
     3421 gtttttatga gggaggcccg aagtcgactt tatcgctgtg acacatactt tctgggcaaa
     3481 acgattgccg aattgccgct ttttctcaca gtgccactgg tcttcacggc gattgcctat
     3541 ccgatgatcg gactgcgggc cggagtgctg cacttcttca actgcctggc gctggtcact
     3601 ctggtggcca atgtgtcaac gtccttcgga tatctaatat cctgcgccag ctcctcgacc
     3661 tcgatggcgc tgtctgtggg tccgccggtt atcataccat tcctgctctt tggcggcttc
     3721 ttcttgaact cgggctcggt gccagtatac ctcaaatggt tgtcgtacct ctcatggttc
     3781 cgttacgcca acgagggtct gctgattaac caatgggcgg acgtggagcc gggcgaaatt
     3841 agctgcacat cgtcgaacac cacgtgcccc agttcgggca aggtcatcct ggagacgctt
     3901 aacttctccg ccgccgatct gccgctggac tacgtgggtc tggccattct catcgtgagc
     3961 ttccgggtgc tcgcatatct ggctctaaga cttcgggccc gacgcaagga gtagccgaca
     4021 tatatccgaa ataactgctt gttttttttt tttaccatta ttaccatcgt gtttactgtt
     4081 tattgccccc tcaaaaagct aatgtaatta tatttgtgcc aataaaaaca agatatgacc
     4141 tatagaatac aagtatttcc ccttcgaaca tccccacaag tagactttgg atttgtcttc
     4201 taaccaaaag acttacacac ctgcatacct tacatcaaaa actcgtttat cgctacataa
     4261 aacaccggga tatatttttt atatacatac ttttcaaatc gcgcgccctc ttcataattc
     4321 acctccacca caccacgttt cgtagttgct ctttcgctgt ctcccacccg ctctccgcaa
     4381 cacattcacc ttttgttcga cgaccttgga gcgactgtcg ttagttccgc gcgattcggt
     4441 tcgctcaaat ggttccgagt ggttcatttc gtctcaatag aaattagtaa taaatatttg
     4501 tatgtacaat ttatttgctc caatatattt gtatatattt ccctcacagc tatatttatt
     4561 ctaatttaat attatgactt tttaaggtaa ttttttgtga cctgttcgga gtgattagcg
     4621 ttacaatttg aactgaaagt gacatccagt gtttgttcct tgtgtagatg catctcaaaa
     4681 aaatggtggg cataatagtg ttgtttatat atatcaaaaa taacaactat aataataaga
     4741 atacatttaa tttagaaaat gcttggattt cactggaact aggctagcat aacttcgtat
     4801 aatgtatgct atacgaagtt atgctagcgg atccgggaat tgggaattgg gcaatattta
     4861 aatggcggcg cgccgaattc ttttttgctc acctgtgatt gctcctactc aaatacaaaa
     4921 acatcaaatt ttctgtcaat aaagcatatt tatttatatt tattttacag gaaagaattc
     4981 cttttaaagt gtattttaac ctataatgaa aaacgattaa aaaaaataca taaaataatt
     5041 cgaaaatttt tgaatagccc aggttgataa aaattcattt catacgtttt ataacttatg
     5101 cccctaagta ttttttgacc atagtgtttc aattctacat taattttaca gagtagaatg
     5161 aaacgccacc tactcagcca agaggcgaaa aggttagctc gccaagcaga gagggcgcca
     5221 gtgctcacta ctttttataa ttctcaactt ctttttccag actcagttcg tatatataga
     5281 cctattttca atttaacgtc gcggcgagaa agagaatacc agttttagag ctagaaatag
     5341 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     5401 ttttttttgg tacctctaga gatccactag tgtcgacgat gtaggtcacg gtctcgaagc
     5461 cgcggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc acctcaccca
     5521 tctggtccat catgatgaac gggtcgaggt ggcggtagtt gatcccggcg aacgcgcggc
     5581 gcaccgggaa gccctcgccc tcgaaaccgc tgggcgcggt ggtcacggtg agcacgggac
     5641 gtgcgacggc gtcggcgggt gcggatacgc ggggcagcgt cagcgggttc tcgacggtca
     5701 cggcgggcat gtcgacacta gttctagcca gcttttgttc cctttagtga gggttaattt
     5761 cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5821 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5881 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5941 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6001 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6061 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7921 aagtgccac
//
