LOCUS       sequence-151054-        6582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  QF2 HACK backbone compatible with GMR attP integrated insertions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  AUTHORS   Lin CC, Potter CJ
  TITLE     Editing Transgenic DNA Components by Inducible Gene Replacement in 
            Drosophila melanogaster.
  JOURNAL   Genetics. 2016 Jun 22. pii: genetics.116.191783.
  PUBMED    27334272
REFERENCE   2  (bases 1 to 6582)
  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: "Genetics. 
            2016 Jun 22. pii: genetics.116.191783."
COMMENT     Sequence Label: pBPGUw-HACK-QF2
FEATURES             Location/Qualifiers
     source          1..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(141..729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(221..240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(900..1691,1692..1760))
                     /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     1523..1542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1761..1865)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1933..1951
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2008..2061
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(3394..3427)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3491..3612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3521)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3556..3575
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3732..4442)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4279..4297
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(4620..4653)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4676..5472
                     /label=dU6-1 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-1 
                     snRNA (Port et al., 2014)"
     misc_RNA        5491..5566
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5567..5987
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     misc_RNA        6006..6081
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    join(6528..6582,1..15)
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 caccgcggct tcgagaccgt gacctacatc gtcgacggta acatgtgaag tttctatact
       61 ttctagagaa tagaaacttc catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      121 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      181 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      241 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      301 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      361 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      481 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      541 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      601 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      661 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      721 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      781 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      841 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
      901 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
      961 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1021 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1081 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1141 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1201 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1261 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1321 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1381 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1441 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1501 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1561 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1621 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1681 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1741 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1801 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1861 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     1921 ttaacctata aaaataggcg tatcacgagg ccctttcgta cgaagtttct atactttcta
     1981 gagaatagaa acttctacgt aacgcgtgag ggccgcggca gcctgctgac ctgcggcgat
     2041 gtggaggaga accccgggcc ccctaggatg ccacccaagc gcaaaacgct taacgctgcg
     2101 gctgaggcta acgctcatgc cgacggacac gccgacggaa acgccgacgg acacgtggcc
     2161 aatacggccg cgtcctcgaa taatgcgagg ttcgctgatc tcactaacat cgatactccg
     2221 ggtctgggac ccacaactac gaccctgctc gtggaaccag cacgctcaaa gcgtcaacga
     2281 gtgtcccgcg catgcgacca gtgccgtgca gcccgagaga aatgcgacgg aatacagcct
     2341 gcgtgtttcc cgtgcgtttc ccagggaagg tcctgcactt atcaggcttc gccgaaaaag
     2401 aggggagttc aaaccggtta tattcgtacg ctggagctcg ccctcgcctg gatgtttgaa
     2461 aatgtcgcgc gttccgaaga tgccttgcat aacctcctcg tccgtgacgc cggacaagga
     2521 tcagctctgc tcgttggtaa agattcgccg gctgccgagc gactccatgc ccgttgggct
     2581 actagccgtg tcaataagag cattacccgc ctcctccgtc agttggagct ccctcctacc
     2641 gccacggcta cggcctcgat aatgccgcac gtgatggagc agcctctcag taccagcatt
     2701 aaccccgtca acgaccgctt caacggtatt cccaacccca ctccgtataa ctccgatgca
     2761 gctctcgatg ctatcactca gaccaacgat tatggaagcg taaatacaca tggtatcctc
     2821 tctacttacc cgccaccggc tacgcacctt aatgaagctt ccgtcgctct cgctcccggt
     2881 ggcgcccccc cccgaccgcc tcctccgtat gttgacagca cgaccaatca cccgccgtac
     2941 cactcgaatc tggttccaat ggcgaacttt ggttactcga ccgttgatta cgatgccatg
     3001 gttgacgatt tggctagcat tgaatacacg gacgctgtgg atgtcgaccc acagtttatg
     3061 accaatctgg gattcgttcc tggatgtaac ttctccgaca ttaatacata cgaacagtga
     3121 tgacgtctat cgataccgtc gactaaagcc aaatagaaaa ttattcagtt cctggcttaa
     3181 gtttttaaaa gtgatattat ttatttggtt gtaaccaacc aaaagaatgt aaataactaa
     3241 tacataatta tgttagtttt aagttagcaa caaattgatt ttagctatat tagctacttg
     3301 gttaataaat agaatatatt tatttaaaga taattgcgtt tttattgtca gggagtgagt
     3361 ttgcttaaaa actcgtttag atccaccgca tgcataactt cgtatagcat acattatacg
     3421 aagttatggt accactcgac gattaaccgt tagtttagca agtataattg aattttttat
     3481 aagatccagc taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3541 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3601 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3661 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag
     3721 agtcgcggcg cttacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg
     3781 cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg
     3841 ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc
     3901 tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg
     3961 gcgccgtcct cggggtacat ccgctcggag gaggcctccc agcccatggt tttcttctgc
     4021 attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg
     4081 ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc
     4141 acgcgctccc acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg
     4201 gggtgcttca cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg
     4261 aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg
     4321 cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc
     4381 accttgaagc gcatgaactc cttgatgatg gccatgttat cctcctcgcc cttgctcacc
     4441 atgggccgat tgtttagctt gttcagctgc gcttgtttat ttgcttagct ttcgcttagc
     4501 gacgtgttca ctttgcttgt ttgaattgaa ttgtcgctcc gtagacgaag cgcctctatt
     4561 tatactccgg cgctaattga attagctcta attgaattag tctctaattg aatgacgtca
     4621 taacttcgta tagcatacat tatacgaagt tatagatcac tagtgatctg aattcatttt
     4681 caacgtcctc gatagtatag tggttagtat ccccgcctgt cacgcgggag accggggttc
     4741 aattccccgt cggggagaat ctgtgattct tttttttttt cttttacttt gttatataaa
     4801 caatttttgt tttaattgaa tctaatttgc cattgctttt aggaatctca ggcatccagc
     4861 aagcgtttgt ccgccgaatc gcccatcagt gaagaagatc ctgtggcggc tacgaaaatc
     4921 tccccggcca tgtcggcctc cacctccagc gaaaaaccca tcagcgagct ggccacctct
     4981 gtgctgaccc accgctttcc agactccacc tcctcacccg gcgaacatgg ccttggacga
     5041 atgcagttgt cgatccgcta cagcgcccag cgtcaaaaac tagacgtgac catacacaaa
     5101 atccagaaga taccacttcg cgatcccagc aatatccccg atccgtatgt taagctgtat
     5161 ctgttgcctg gacgcaccaa ggagtcgaaa cgcaagacga gcgtgatcaa ggacaactgc
     5221 aaccccgtct acgatgcatc ctttgagtac ctgatttcca ttgccgaact caggcagacg
     5281 gaactggagg tgacggtgtg cacccaaaag ggattcctat ccggcggtag tcccatcatt
     5341 ggcatggtag gtacccgaaa gcaacccctt agttacagac acagcgcgta cgtccttcgc
     5401 atccttatga ttcccaagta catattctgc aagagtacag tatatatagg aaagatatcc
     5461 gggtgaactt cggggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat
     5521 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgctcacct
     5581 gtgattgctc ctactcaaat acaaaaacat caaattttct gtcaataaag catatttatt
     5641 tatatttatt ttacaggaaa gaattccttt taaagtgtat tttaacctat aatgaaaaac
     5701 gattaaaaaa aatacataaa ataattcgaa aatttttgaa tagcccaggt tgataaaaat
     5761 tcatttcata cgttttataa cttatgcccc taagtatttt ttgaccatag tgtttcaatt
     5821 ctacattaat tttacagagt agaatgaaac gccacctact cagccaagag gcgaaaaggt
     5881 tagctcgcca agcagagagg gcgccagtgc tcactacttt ttataattct caacttcttt
     5941 ttccagactc agttcgtata tatagaccta ttttcaattt aacgtcgggg tcttcgagaa
     6001 gacctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     6061 aaagtggcac cgagtcggtg cttttttgcc tacctggagc ctgagagttg ttcaataaaa
     6121 taaaaatgtt tcgttttttt gctttcgcca gtatttatta tttttcatca atatgtattc
     6181 aatttggtat gtatttagta attgtaatat atagacaatg gttttccgtt gacgtacata
     6241 catctgacgt gtgtttattt agacataata gttatgtttt cacatctttt taatgttcgc
     6301 ttaatgcgta tgcttaatta atatgttcgg cttgtcgaca tgcccgccgt gaccgtcgag
     6361 aacccgctga cgctgccccg cgtatccgca cccgccgacg ccgtcgcacg tcccgtgctc
     6421 accgtgacca ccgcgcccag cggtttcgag ggcgagggct tcccggtgcg ccgcgcgttc
     6481 gccgggatca actaccgcca cctcgacccg ttcatcatga tggaccagat gggtgaggtg
     6541 gagtacgcgc ccggggagcc caagggcacg ccctggcacc cg
//
