LOCUS       sequence-374378-        8373 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 8373)
  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 8373)
  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: p1X-UASTattB
FEATURES             Location/Qualifiers
     source          1..8373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(32..50)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(71..87)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(71..87)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(84..106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    95..111
                     /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(119..125,126..143,144..149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    164..185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(302..319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(473..1061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(553..572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1232..2023,2024..2092))
                     /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     1855..1874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2093..2197)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2223..2678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2360..2381)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2572..2591
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2805..2827
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2819..2836
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2820..2836
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2843..2862
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2843..2861
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2895..2913)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(3365..3436,3838..4111,4186..4840,4902..5217,5421..5552,
                     5623..6237)
                     /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    7012..7045
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7101..7339
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     7286..7307
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          7467..7532
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7662..7682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7954..8088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7991..8010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8045..8064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    8117..8186
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 tgtcgacact agttctagcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
       61 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     1861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     1921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2221 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2281 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2341 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2401 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2461 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2521 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2581 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2641 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     2701 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2761 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2821 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
     2881 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
     2941 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
     3001 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
     3061 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
     3121 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
     3181 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     3241 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     3301 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     3361 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     3421 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     3481 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     3541 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     3601 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     3661 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     3721 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     3781 cggggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     3841 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     3901 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     3961 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     4021 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     4081 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     4141 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     4201 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     4261 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     4321 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     4381 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccaa gccatggtgc
     4441 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     4501 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     4561 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggctctaacc gatccgccgc
     4621 ttctgatctg cgatgagccc acctccggac tggactcctt taccgcccac agcgtcgtcc
     4681 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     4741 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     4801 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     4861 aagggtatct agtattacat aacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     4921 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     4981 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgccattag caaagtagcc
     5041 cgggatatgg agcagttgtt ggccaccaaa aatctggaga agccactgga gcagccggag
     5101 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     5161 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     5221 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     5281 attttagata gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     5341 tttatagttt tgcagaagat aaataaattt catttaactc gcgaatatta atgagatgcg
     5401 agtaacattt taatttgcag atggttgcca tcttgattgg cctcatcttt ttgggccaac
     5461 aactcacgca agtgggtgtg atgaatatca acggagccat cttcctcttc ctgaccaaca
     5521 tgacctttca aaacgtcttt gccacgataa atgtaagtca tgtttagaat acatttgcat
     5581 ttcaataatt tactaacttt ctaatgaatc gattcgattt aggtgttcac ctcagagctg
     5641 ccagttttta tgagggaggc ccgaagtcga ctttatcgct gtgacacata ctttctgggc
     5701 aaaacgattg ccgaattgcc gctttttctc acagtgccac tggtcttcac ggcgattgcc
     5761 tatccgatga tcggactgcg ggccggagtg ctgcacttct tcaactgcct ggcgctggtc
     5821 actctggtgg ccaatgtgtc aacgtccttc ggatatctaa tatcctgcgc cagctcctcg
     5881 acctcgatgg cgctgtctgt gggtccgccg gttatcatac cattcctgct ctttggcggc
     5941 ttcttcttga actcgggctc ggtgccagta tacctcaaat ggttgtcgta cctctcatgg
     6001 ttccgttacg ccaacgaggg tctgctgatt aaccaatggg cggacgtgga gccgggcgaa
     6061 attagctgca catcgtcgaa caccacgtgc cccagttcgg gcaaggtcat cctggagacg
     6121 cttaacttct ccgccgccga tctgccgctg gactacgtgg gtctggccat tctcatcgtg
     6181 agcttccggg tgctcgcata tctggctcta agacttcggg cccgacgcaa ggagtagccg
     6241 acatatatcc gaaataactg cttgtttttt ttttttacca ttattaccat cgtgtttact
     6301 gtttattgcc ccctcaaaaa gctaatgtaa ttatatttgt gccaataaaa acaagatatg
     6361 acctatagaa tacaagtatt tccccttcga acatccccac aagtagactt tggatttgtc
     6421 ttctaaccaa aagacttaca cacctgcata ccttacatca aaaactcgtt tatcgctaca
     6481 taaaacaccg ggatatattt tttatataca tacttttcaa atcgcgcgcc ctcttcataa
     6541 ttcacctcca ccacaccacg tttcgtagtt gctctttcgc tgtctcccac ccgctctccg
     6601 caacacattc accttttgtt cgacgacctt ggagcgactg tcgttagttc cgcgcgattc
     6661 ggttcgctca aatggttccg agtggttcat ttcgtctcaa tagaaattag taataaatat
     6721 ttgtatgtac aatttatttg ctccaatata tttgtatata tttccctcac agctatattt
     6781 attctaattt aatattatga ctttttaagg taattttttg tgacctgttc ggagtgatta
     6841 gcgttacaat ttgaactgaa agtgacatcc agtgtttgtt ccttgtgtag atgcatctca
     6901 aaaaaatggt gggcataata gtgttgttta tatatatcaa aaataacaac tataataata
     6961 agaatacatt taatttagaa aatgcttgga tttcactgga actaggctag cataacttcg
     7021 tataatgtat gctatacgaa gttatgctag cggatccaag cttcggagta ctgtcctccg
     7081 agcggagact ctagcgagcg ccggagtata aatagaggcg cttcgtctac ggagcgacaa
     7141 ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc
     7201 gcagctgaac aagctaaaca atctgcagta aagtgcaagt taaagtgaat caattaaaag
     7261 taaccagcaa ccaagtaaat caactgcaac tactgaaatc tgccaagaag taattattga
     7321 atacaagaag agaactctga atagaattcg ttaacagatc tgcggccgcg gctcgagggt
     7381 acctctagag gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac
     7441 tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta
     7501 aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact gatgaatggg
     7561 agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa atgccatcta
     7621 gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag aagagaaagg
     7681 tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat gctgtgttta
     7741 gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct gcactgctat
     7801 acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat cataatcagc
     7861 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     7921 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     7981 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     8041 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcca ctagtgtcga
     8101 cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc ttgggctccc
     8161 cgggcgcgta ctccacctca cccatctggt ccatcatgat gaacgggtcg aggtggcggt
     8221 agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc gccctcgaaa ccgctgggcg
     8281 cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc gggtgcggat acgcggggca
     8341 gcgtcagcgg gttctcgacg gtcacggcgg gca
//
