LOCUS       sequence-374376-        8430 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 8430)
  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 8430)
  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: p4X-UASTattB
FEATURES             Location/Qualifiers
     source          1..8430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..61
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          221..286
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             416..436
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    708..842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(745..764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     799..818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    871..940
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(1159..1177)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1198..1214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1198..1214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1211..1233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1222..1238
                     /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(1246..1252,1253..1270,1271..1276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1291..1312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1429..1446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1600..2188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1680..1699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2359..3150,3151..3219))
                     /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     2982..3001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3220..3324)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3350..3805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3487..3508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3699..3718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3932..3954
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3946..3963
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3947..3963
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3970..3989
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3970..3988
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4022..4040)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(4492..4563,4965..5238,5313..5967,6029..6344,6548..6679,
                     6750..7364)
                     /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    8139..8172
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8285..8430,1..93)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
ORIGIN
        1 gaatcaatta aaagtaacca gcaaccaagt aaatcaactg caactactga aatctgccaa
       61 gaagtaatta ttgaatacaa gaagagaact ctgaatagaa ttcgttaaca gatctgcggc
      121 cgcggctcga gggtacctct agaggatctt tgtgaaggaa ccttacttct gtggtgtgac
      181 ataattggac aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag
      241 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg
      301 aactgatgaa tgggagcagt ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga
      361 agaaatgcca tctagtgatg atgaggctac tgctgactct caacattcta ctcctccaaa
      421 aaagaagaga aaggtagaag accccaagga ctttccttca gaattgctaa gttttttgag
      481 tcatgctgtg tttagtaata gaactcttgc ttgctttgct atttacacca caaaggaaaa
      541 agctgcactg ctatacaaga aaattatgga aaaatatttg atgtatagtg ccttgactag
      601 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
      661 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
      721 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
      781 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
      841 tccactagtg tcgacgatgt aggtcacggt ctcgaagccg cggtgcgggt gccagggcgt
      901 gcccttgggc tccccgggcg cgtactccac ctcacccatc tggtccatca tgatgaacgg
      961 gtcgaggtgg cggtagttga tcccggcgaa cgcgcggcgc accgggaagc cctcgccctc
     1021 gaaaccgctg ggcgcggtgg tcacggtgag cacgggacgt gcgacggcgt cggcgggtgc
     1081 ggatacgcgg ggcagcgtca gcgggttctc gacggtcacg gcgggcatgt cgacactagt
     1141 tctagccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
     1201 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1261 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1321 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1381 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1441 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1501 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1561 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1621 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1681 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1741 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1801 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1861 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1921 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1981 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2041 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2101 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2161 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2221 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2281 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2401 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2461 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2521 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2581 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2641 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2701 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2761 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2821 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2881 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2941 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3001 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3061 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3121 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3181 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3241 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3301 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     3361 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3421 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3481 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3541 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3601 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3661 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3721 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3781 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     3841 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     3901 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     3961 agtgaattgt aatacgactc actatagggc gaattgggta cgtaccgggc ccctagtatg
     4021 tatgtaagtt aataaaaccc atttttgcgg aaagtagata aaaaaaacat tttttttttt
     4081 tactgcactg gatatcattg aacttatctg atcagtttta aatttacttc gatccaaggg
     4141 tatttgatgt accaggttct ttcgattacc tctcactcaa aatgacattc cactcaaagt
     4201 cagcgctgtt tgcctccttc tctgtccaca gaaatatcgc cgtctctttc gccgctgcgt
     4261 ccgctatctc tttcgccacc gtttgtagcg ttacgtagcg tcaatgtccg ccttcagttg
     4321 cattttgtca gcggtttcgt gacgaagctc caagcggttt acgccatcaa ttaaacacaa
     4381 agtgctgtgc caaaactcct ctcgcttctt atttttgttt gttttttgag tgattggggt
     4441 ggtgattggt tttgggtggg taagcagggg aaagtgtgaa aaatcccggc aatgggccaa
     4501 gaggatcagg agctattaat tcgcggaggc agcaaacacc catctgccga gcatctgaac
     4561 aatgtgagta gtacatgtgc atacatctta agttcacttg atctatagga actgcgattg
     4621 caacatcaaa ttgtctgcgg cgtgagaact gcgacccaca aaaatcccaa accgcaattg
     4681 cacaaacaaa tagtgacacg aaacagatta ttctggtagc tgttctcgct atataagaca
     4741 atttttgaga tcatatcatg atcaagacat ctaaaggcat tcattttcga ctatattctt
     4801 ttttacaaaa aatataacaa ccagatattt taagctgatc ctagatgcac aaaaaataaa
     4861 taaaagtata aacctacttc gtaggatact tcggggtact ttttgttcgg ggttagatga
     4921 gcataacgct tgtagttgat atttgagatc ccctatcatt gcagggtgac agcggagcgg
     4981 cttcgcagag ctgcattaac cagggcttcg ggcaggccaa aaactacggc acgctccggc
     5041 cacccagtcc gccggaggac tccggttcag ggagcggcca actagccgag aacctcacct
     5101 atgcctggca caatatggac atctttgggg cggtcaatca gccgggctcc ggatggcggc
     5161 agctggtcaa ccggacacgc ggactattct gcaacgagcg acacataccg gcgcccagga
     5221 aacatttgct caagaacggt gagtttctat tcgcagtcgg ctgatctgtg tgaaatctta
     5281 ataaagggtc caattaccaa tttgaaactc agtttgcggc gtggcctatc cgggcgaact
     5341 tttggccgtg atgggcagtt ccggtgccgg aaagacgacc ctgctgaatg cccttgcctt
     5401 tcgatcgccg cagggcatcc aagtatcgcc atccgggatg cgactgctca atggccaacc
     5461 tgtggacgcc aaggagatgc aggccaggtg cgcctatgtc cagcaggatg acctctttat
     5521 cggctcccta acggccaggg aacacctgat tttccaagcc atggtgcgga tgccacgaca
     5581 tctgacctat cggcagcgag tggcccgcgt ggatcaggtg atccaggagc tttcgctcag
     5641 caaatgtcag cacacgatca tcggtgtgcc cggcagggtg aaaggtctgt ccggcggaga
     5701 aaggaagcgt ctggcattcg cctccgaggc tctaaccgat ccgccgcttc tgatctgcga
     5761 tgagcccacc tccggactgg actcctttac cgcccacagc gtcgtccagg tgctgaagaa
     5821 gctgtcgcag aagggcaaga ccgtcatcct gaccattcat cagccgtctt ccgagctgtt
     5881 tgagctcttt gacaagatcc ttctgatggc cgagggcagg gtagctttct tgggcactcc
     5941 cagcgaagcc gtcgacttct tttcctagtg agttcgatgt gtttattaag ggtatctagt
     6001 attacataac atctcaactc ctatccagcg tgggtgccca gtgtcctacc aactacaatc
     6061 cggcggactt ttacgtacag gtgttggccg ttgtgcccgg acgggagatc gagtcccgtg
     6121 atcggatcgc caagatatgc gacaattttg ccattagcaa agtagcccgg gatatggagc
     6181 agttgttggc caccaaaaat ctggagaagc cactggagca gccggagaat gggtacacct
     6241 acaaggccac ctggttcatg cagttccggg cggtcctgtg gcgatcctgg ctgtcggtgc
     6301 tcaaggaacc actcctcgta aaagtgcgac ttattcagac aacggtgagt ggttccagtg
     6361 gaaacaaatg atataacgct tacaattctt ggaaacaaat tcgctagatt ttagatagaa
     6421 ttgcctgatt ccacaccctt cttagttttt ttcaatgaga tgtatagttt atagttttgc
     6481 agaagataaa taaatttcat ttaactcgcg aatattaatg agatgcgagt aacattttaa
     6541 tttgcagatg gttgccatct tgattggcct catctttttg ggccaacaac tcacgcaagt
     6601 gggtgtgatg aatatcaacg gagccatctt cctcttcctg accaacatga cctttcaaaa
     6661 cgtctttgcc acgataaatg taagtcatgt ttagaataca tttgcatttc aataatttac
     6721 taactttcta atgaatcgat tcgatttagg tgttcacctc agagctgcca gtttttatga
     6781 gggaggcccg aagtcgactt tatcgctgtg acacatactt tctgggcaaa acgattgccg
     6841 aattgccgct ttttctcaca gtgccactgg tcttcacggc gattgcctat ccgatgatcg
     6901 gactgcgggc cggagtgctg cacttcttca actgcctggc gctggtcact ctggtggcca
     6961 atgtgtcaac gtccttcgga tatctaatat cctgcgccag ctcctcgacc tcgatggcgc
     7021 tgtctgtggg tccgccggtt atcataccat tcctgctctt tggcggcttc ttcttgaact
     7081 cgggctcggt gccagtatac ctcaaatggt tgtcgtacct ctcatggttc cgttacgcca
     7141 acgagggtct gctgattaac caatgggcgg acgtggagcc gggcgaaatt agctgcacat
     7201 cgtcgaacac cacgtgcccc agttcgggca aggtcatcct ggagacgctt aacttctccg
     7261 ccgccgatct gccgctggac tacgtgggtc tggccattct catcgtgagc ttccgggtgc
     7321 tcgcatatct ggctctaaga cttcgggccc gacgcaagga gtagccgaca tatatccgaa
     7381 ataactgctt gttttttttt tttaccatta ttaccatcgt gtttactgtt tattgccccc
     7441 tcaaaaagct aatgtaatta tatttgtgcc aataaaaaca agatatgacc tatagaatac
     7501 aagtatttcc ccttcgaaca tccccacaag tagactttgg atttgtcttc taaccaaaag
     7561 acttacacac ctgcatacct tacatcaaaa actcgtttat cgctacataa aacaccggga
     7621 tatatttttt atatacatac ttttcaaatc gcgcgccctc ttcataattc acctccacca
     7681 caccacgttt cgtagttgct ctttcgctgt ctcccacccg ctctccgcaa cacattcacc
     7741 ttttgttcga cgaccttgga gcgactgtcg ttagttccgc gcgattcggt tcgctcaaat
     7801 ggttccgagt ggttcatttc gtctcaatag aaattagtaa taaatatttg tatgtacaat
     7861 ttatttgctc caatatattt gtatatattt ccctcacagc tatatttatt ctaatttaat
     7921 attatgactt tttaaggtaa ttttttgtga cctgttcgga gtgattagcg ttacaatttg
     7981 aactgaaagt gacatccagt gtttgttcct tgtgtagatg catctcaaaa aaatggtggg
     8041 cataatagtg ttgtttatat atatcaaaaa taacaactat aataataaga atacatttaa
     8101 tttagaaaat gcttggattt cactggaact aggctagcat aacttcgtat aatgtatgct
     8161 atacgaagtt atgctagcgg atccaagctt cggagtactg tcctccgagc ggagtactgt
     8221 cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gactctagcg
     8281 agcgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc
     8341 aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct gaacaagcta
     8401 aacaatctgc agtaaagtgc aagttaaagt
//
