LOCUS       sequence-374377-        8392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fly cloning and transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8392)
  AUTHORS   Singh M, Kim JH
  TITLE     Generation and validation of pX-UASTattB for dose-dependent 
            misexpression studies in Drosophila.
  JOURNAL   MicroPubl Biol. 2022 Aug 12;2022. doi: 
            10.17912/micropub.biology.000626. eCollection 2022.
  PUBMED    36039330
REFERENCE   2  (bases 1 to 8392)
  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: "MicroPubl 
            Biol."; date: "2022-08-12"; volume: "2022. doi"; pages: " 
            10.17912/micropub.biology.000626. eCollection 2022"
COMMENT     Sequence Label: p2X-UASTattB
FEATURES             Location/Qualifiers
     source          1..8392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..31
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    84..153
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(372..390)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(411..427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(411..427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(424..446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    435..451
                     /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(459..465,466..483,484..489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    504..525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(642..659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(813..1401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(893..912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1572..2363,2364..2432))
                     /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     2195..2214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2433..2537)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2563..3018)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2700..2721)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2912..2931
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3145..3167
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3159..3176
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3160..3176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3183..3202
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3183..3201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3235..3253)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(3705..3776,4178..4451,4526..5180,5242..5557,5761..5892,
                     5963..6577)
                     /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    7352..7385
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7460..7698
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     7645..7666
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          7826..7891
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8021..8041
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    join(8313..8392,1..55)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8350..8369)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccacta
       61 gtgtcgacga tgtaggtcac ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg
      121 ggctccccgg gcgcgtactc cacctcaccc atctggtcca tcatgatgaa cgggtcgagg
      181 tggcggtagt tgatcccggc gaacgcgcgg cgcaccggga agccctcgcc ctcgaaaccg
      241 ctgggcgcgg tggtcacggt gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg
      301 cggggcagcg tcagcgggtt ctcgacggtc acggcgggca tgtcgacact agttctagcc
      361 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
      421 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      481 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      541 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      601 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      661 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      721 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      781 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      841 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      901 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      961 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1021 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1081 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1141 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1201 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1261 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1321 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1381 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1441 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1501 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1561 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1621 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1681 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1741 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1801 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1861 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1921 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1981 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2041 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2101 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2161 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2221 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2281 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2341 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2401 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2461 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2521 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     2581 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2641 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2701 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2761 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2821 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2881 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2941 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3001 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     3061 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     3121 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     3181 tgtaatacga ctcactatag ggcgaattgg gtacgtaccg ggcccctagt atgtatgtaa
     3241 gttaataaaa cccatttttg cggaaagtag ataaaaaaaa catttttttt ttttactgca
     3301 ctggatatca ttgaacttat ctgatcagtt ttaaatttac ttcgatccaa gggtatttga
     3361 tgtaccaggt tctttcgatt acctctcact caaaatgaca ttccactcaa agtcagcgct
     3421 gtttgcctcc ttctctgtcc acagaaatat cgccgtctct ttcgccgctg cgtccgctat
     3481 ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt ccgccttcag ttgcattttg
     3541 tcagcggttt cgtgacgaag ctccaagcgg tttacgccat caattaaaca caaagtgctg
     3601 tgccaaaact cctctcgctt cttatttttg tttgtttttt gagtgattgg ggtggtgatt
     3661 ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc ggcaatgggc caagaggatc
     3721 aggagctatt aattcgcgga ggcagcaaac acccatctgc cgagcatctg aacaatgtga
     3781 gtagtacatg tgcatacatc ttaagttcac ttgatctata ggaactgcga ttgcaacatc
     3841 aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc caaaccgcaa ttgcacaaac
     3901 aaatagtgac acgaaacaga ttattctggt agctgttctc gctatataag acaatttttg
     3961 agatcatatc atgatcaaga catctaaagg cattcatttt cgactatatt cttttttaca
     4021 aaaaatataa caaccagata ttttaagctg atcctagatg cacaaaaaat aaataaaagt
     4081 ataaacctac ttcgtaggat acttcggggt actttttgtt cggggttaga tgagcataac
     4141 gcttgtagtt gatatttgag atcccctatc attgcagggt gacagcggag cggcttcgca
     4201 gagctgcatt aaccagggct tcgggcaggc caaaaactac ggcacgctcc ggccacccag
     4261 tccgccggag gactccggtt cagggagcgg ccaactagcc gagaacctca cctatgcctg
     4321 gcacaatatg gacatctttg gggcggtcaa tcagccgggc tccggatggc ggcagctggt
     4381 caaccggaca cgcggactat tctgcaacga gcgacacata ccggcgccca ggaaacattt
     4441 gctcaagaac ggtgagtttc tattcgcagt cggctgatct gtgtgaaatc ttaataaagg
     4501 gtccaattac caatttgaaa ctcagtttgc ggcgtggcct atccgggcga acttttggcc
     4561 gtgatgggca gttccggtgc cggaaagacg accctgctga atgcccttgc ctttcgatcg
     4621 ccgcagggca tccaagtatc gccatccggg atgcgactgc tcaatggcca acctgtggac
     4681 gccaaggaga tgcaggccag gtgcgcctat gtccagcagg atgacctctt tatcggctcc
     4741 ctaacggcca gggaacacct gattttccaa gccatggtgc ggatgccacg acatctgacc
     4801 tatcggcagc gagtggcccg cgtggatcag gtgatccagg agctttcgct cagcaaatgt
     4861 cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc tgtccggcgg agaaaggaag
     4921 cgtctggcat tcgcctccga ggctctaacc gatccgccgc ttctgatctg cgatgagccc
     4981 acctccggac tggactcctt taccgcccac agcgtcgtcc aggtgctgaa gaagctgtcg
     5041 cagaagggca agaccgtcat cctgaccatt catcagccgt cttccgagct gtttgagctc
     5101 tttgacaaga tccttctgat ggccgagggc agggtagctt tcttgggcac tcccagcgaa
     5161 gccgtcgact tcttttccta gtgagttcga tgtgtttatt aagggtatct agtattacat
     5221 aacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct accaactaca atccggcgga
     5281 cttttacgta caggtgttgg ccgttgtgcc cggacgggag atcgagtccc gtgatcggat
     5341 cgccaagata tgcgacaatt ttgccattag caaagtagcc cgggatatgg agcagttgtt
     5401 ggccaccaaa aatctggaga agccactgga gcagccggag aatgggtaca cctacaaggc
     5461 cacctggttc atgcagttcc gggcggtcct gtggcgatcc tggctgtcgg tgctcaagga
     5521 accactcctc gtaaaagtgc gacttattca gacaacggtg agtggttcca gtggaaacaa
     5581 atgatataac gcttacaatt cttggaaaca aattcgctag attttagata gaattgcctg
     5641 attccacacc cttcttagtt tttttcaatg agatgtatag tttatagttt tgcagaagat
     5701 aaataaattt catttaactc gcgaatatta atgagatgcg agtaacattt taatttgcag
     5761 atggttgcca tcttgattgg cctcatcttt ttgggccaac aactcacgca agtgggtgtg
     5821 atgaatatca acggagccat cttcctcttc ctgaccaaca tgacctttca aaacgtcttt
     5881 gccacgataa atgtaagtca tgtttagaat acatttgcat ttcaataatt tactaacttt
     5941 ctaatgaatc gattcgattt aggtgttcac ctcagagctg ccagttttta tgagggaggc
     6001 ccgaagtcga ctttatcgct gtgacacata ctttctgggc aaaacgattg ccgaattgcc
     6061 gctttttctc acagtgccac tggtcttcac ggcgattgcc tatccgatga tcggactgcg
     6121 ggccggagtg ctgcacttct tcaactgcct ggcgctggtc actctggtgg ccaatgtgtc
     6181 aacgtccttc ggatatctaa tatcctgcgc cagctcctcg acctcgatgg cgctgtctgt
     6241 gggtccgccg gttatcatac cattcctgct ctttggcggc ttcttcttga actcgggctc
     6301 ggtgccagta tacctcaaat ggttgtcgta cctctcatgg ttccgttacg ccaacgaggg
     6361 tctgctgatt aaccaatggg cggacgtgga gccgggcgaa attagctgca catcgtcgaa
     6421 caccacgtgc cccagttcgg gcaaggtcat cctggagacg cttaacttct ccgccgccga
     6481 tctgccgctg gactacgtgg gtctggccat tctcatcgtg agcttccggg tgctcgcata
     6541 tctggctcta agacttcggg cccgacgcaa ggagtagccg acatatatcc gaaataactg
     6601 cttgtttttt ttttttacca ttattaccat cgtgtttact gtttattgcc ccctcaaaaa
     6661 gctaatgtaa ttatatttgt gccaataaaa acaagatatg acctatagaa tacaagtatt
     6721 tccccttcga acatccccac aagtagactt tggatttgtc ttctaaccaa aagacttaca
     6781 cacctgcata ccttacatca aaaactcgtt tatcgctaca taaaacaccg ggatatattt
     6841 tttatataca tacttttcaa atcgcgcgcc ctcttcataa ttcacctcca ccacaccacg
     6901 tttcgtagtt gctctttcgc tgtctcccac ccgctctccg caacacattc accttttgtt
     6961 cgacgacctt ggagcgactg tcgttagttc cgcgcgattc ggttcgctca aatggttccg
     7021 agtggttcat ttcgtctcaa tagaaattag taataaatat ttgtatgtac aatttatttg
     7081 ctccaatata tttgtatata tttccctcac agctatattt attctaattt aatattatga
     7141 ctttttaagg taattttttg tgacctgttc ggagtgatta gcgttacaat ttgaactgaa
     7201 agtgacatcc agtgtttgtt ccttgtgtag atgcatctca aaaaaatggt gggcataata
     7261 gtgttgttta tatatatcaa aaataacaac tataataata agaatacatt taatttagaa
     7321 aatgcttgga tttcactgga actaggctag cataacttcg tataatgtat gctatacgaa
     7381 gttatgctag cggatccaag cttcggagta ctgtcctccg agcggagtac tgtcctccga
     7441 gcggagactc tagcgagcgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat
     7501 tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg
     7561 cagctgaaca agctaaacaa tctgcagtaa agtgcaagtt aaagtgaatc aattaaaagt
     7621 aaccagcaac caagtaaatc aactgcaact actgaaatct gccaagaagt aattattgaa
     7681 tacaagaaga gaactctgaa tagaattcgt taacagatct gcggccgcgg ctcgagggta
     7741 cctctagagg atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact
     7801 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa
     7861 actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga
     7921 gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag
     7981 tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt
     8041 agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag
     8101 taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata
     8161 caagaaaatt atggaaaaat atttgatgta tagtgccttg actagagatc ataatcagcc
     8221 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     8281 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     8341 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ct
//
