LOCUS       sequence-374379-        8411 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 8411)
  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 8411)
  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: p3X-UASTattB
FEATURES             Location/Qualifiers
     source          1..8411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     301..318
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    435..456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(471..476,477..494,495..501)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    509..525
                     /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)."
     primer_bind     514..536
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     533..549
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     533..549
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        570..588
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    807..876
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     polyA_signal    905..1039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(929..948)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     983..1002
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1311..1331)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1461..1526
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        complement(1654..1892)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     complement(1686..1707)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    complement(1986..2019)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(2794..3408,3479..3610,3814..4129,4191..4845,
                     4920..5193,5595..5666))
                     /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"
     primer_bind     6118..6136
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     primer_bind     complement(6169..6188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6170..6188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6195..6212)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6195..6211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6204..6226)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6353..6808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6440..6459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6650..6671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6834..6938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6939..7007,7008..7799)
                     /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     complement(7157..7176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7970..8411,1..147)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
       61 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      121 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      181 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      241 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      301 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      361 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      421 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      481 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      541 agctatgacc atgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg
      601 gctagaacta gtgtcgacat gcccgccgtg accgtcgaga acccgctgac gctgccccgc
      661 gtatccgcac ccgccgacgc cgtcgcacgt cccgtgctca ccgtgaccac cgcgcccagc
      721 ggtttcgagg gcgagggctt cccggtgcgc cgcgcgttcg ccgggatcaa ctaccgccac
      781 ctcgacccgt tcatcatgat ggaccagatg ggtgaggtgg agtacgcgcc cggggagccc
      841 aagggcacgc cctggcaccc gcaccgcggc ttcgagaccg tgacctacat cgtcgacact
      901 agtggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      961 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1021 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1081 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     1141 tgatctctag tcaaggcact atacatcaaa tatttttcca taattttctt gtatagcagt
     1201 gcagcttttt cctttgtggt gtaaatagca aagcaagcaa gagttctatt actaaacaca
     1261 gcatgactca aaaaacttag caattctgaa ggaaagtcct tggggtcttc tacctttctc
     1321 ttcttttttg gaggagtaga atgttgagag tcagcagtag cctcatcatc actagatggc
     1381 atttcttctg agcaaaacag gttttcctca ttaaaggcat tccaccactg ctcccattca
     1441 tcagttccat aggttggaat ctaaaataca caaacaatta gaatcagtag tttaacacat
     1501 tatacactta aaaattttat atttacctta gagctttaaa tctctgtagg tagtttgtcc
     1561 aattatgtca caccacagaa gtaaggttcc ttcacaaaga tcctctagag gtaccctcga
     1621 gccgcggccg cagatctgtt aacgaattct attcagagtt ctcttcttgt attcaataat
     1681 tacttcttgg cagatttcag tagttgcagt tgatttactt ggttgctggt tacttttaat
     1741 tgattcactt taacttgcac tttactgcag attgtttagc ttgttcagct gcgcttgttt
     1801 atttgcttag ctttcgctta gcgacgtgtt cactttgctt gtttgaattg aattgtcgct
     1861 ccgtagacga agcgcctcta tttatactcc ggcgctcgct agagtctccg ctcggaggac
     1921 agtactccgc tcggaggaca gtactccgct cggaggacag tactccgaag cttggatccg
     1981 ctagcataac ttcgtatagc atacattata cgaagttatg ctagcctagt tccagtgaaa
     2041 tccaagcatt ttctaaatta aatgtattct tattattata gttgttattt ttgatatata
     2101 taaacaacac tattatgccc accatttttt tgagatgcat ctacacaagg aacaaacact
     2161 ggatgtcact ttcagttcaa attgtaacgc taatcactcc gaacaggtca caaaaaatta
     2221 ccttaaaaag tcataatatt aaattagaat aaatatagct gtgagggaaa tatatacaaa
     2281 tatattggag caaataaatt gtacatacaa atatttatta ctaatttcta ttgagacgaa
     2341 atgaaccact cggaaccatt tgagcgaacc gaatcgcgcg gaactaacga cagtcgctcc
     2401 aaggtcgtcg aacaaaaggt gaatgtgttg cggagagcgg gtgggagaca gcgaaagagc
     2461 aactacgaaa cgtggtgtgg tggaggtgaa ttatgaagag ggcgcgcgat ttgaaaagta
     2521 tgtatataaa aaatatatcc cggtgtttta tgtagcgata aacgagtttt tgatgtaagg
     2581 tatgcaggtg tgtaagtctt ttggttagaa gacaaatcca aagtctactt gtggggatgt
     2641 tcgaagggga aatacttgta ttctataggt catatcttgt ttttattggc acaaatataa
     2701 ttacattagc tttttgaggg ggcaataaac agtaaacacg atggtaataa tggtaaaaaa
     2761 aaaaaacaag cagttatttc ggatatatgt cggctactcc ttgcgtcggg cccgaagtct
     2821 tagagccaga tatgcgagca cccggaagct cacgatgaga atggccagac ccacgtagtc
     2881 cagcggcaga tcggcggcgg agaagttaag cgtctccagg atgaccttgc ccgaactggg
     2941 gcacgtggtg ttcgacgatg tgcagctaat ttcgcccggc tccacgtccg cccattggtt
     3001 aatcagcaga ccctcgttgg cgtaacggaa ccatgagagg tacgacaacc atttgaggta
     3061 tactggcacc gagcccgagt tcaagaagaa gccgccaaag agcaggaatg gtatgataac
     3121 cggcggaccc acagacagcg ccatcgaggt cgaggagctg gcgcaggata ttagatatcc
     3181 gaaggacgtt gacacattgg ccaccagagt gaccagcgcc aggcagttga agaagtgcag
     3241 cactccggcc cgcagtccga tcatcggata ggcaatcgcc gtgaagacca gtggcactgt
     3301 gagaaaaagc ggcaattcgg caatcgtttt gcccagaaag tatgtgtcac agcgataaag
     3361 tcgacttcgg gcctccctca taaaaactgg cagctctgag gtgaacacct aaatcgaatc
     3421 gattcattag aaagttagta aattattgaa atgcaaatgt attctaaaca tgacttacat
     3481 ttatcgtggc aaagacgttt tgaaaggtca tgttggtcag gaagaggaag atggctccgt
     3541 tgatattcat cacacccact tgcgtgagtt gttggcccaa aaagatgagg ccaatcaaga
     3601 tggcaaccat ctgcaaatta aaatgttact cgcatctcat taatattcgc gagttaaatg
     3661 aaatttattt atcttctgca aaactataaa ctatacatct cattgaaaaa aactaagaag
     3721 ggtgtggaat caggcaattc tatctaaaat ctagcgaatt tgtttccaag aattgtaagc
     3781 gttatatcat ttgtttccac tggaaccact caccgttgtc tgaataagtc gcacttttac
     3841 gaggagtggt tccttgagca ccgacagcca ggatcgccac aggaccgccc ggaactgcat
     3901 gaaccaggtg gccttgtagg tgtacccatt ctccggctgc tccagtggct tctccagatt
     3961 tttggtggcc aacaactgct ccatatcccg ggctactttg ctaatggcaa aattgtcgca
     4021 tatcttggcg atccgatcac gggactcgat ctcccgtccg ggcacaacgg ccaacacctg
     4081 tacgtaaaag tccgccggat tgtagttggt aggacactgg gcacccacgc tggataggag
     4141 ttgagatgtt atgtaatact agataccctt aataaacaca tcgaactcac taggaaaaga
     4201 agtcgacggc ttcgctggga gtgcccaaga aagctaccct gccctcggcc atcagaagga
     4261 tcttgtcaaa gagctcaaac agctcggaag acggctgatg aatggtcagg atgacggtct
     4321 tgcccttctg cgacagcttc ttcagcacct ggacgacgct gtgggcggta aaggagtcca
     4381 gtccggaggt gggctcatcg cagatcagaa gcggcggatc ggttagagcc tcggaggcga
     4441 atgccagacg cttcctttct ccgccggaca gacctttcac cctgccgggc acaccgatga
     4501 tcgtgtgctg acatttgctg agcgaaagct cctggatcac ctgatccacg cgggccactc
     4561 gctgccgata ggtcagatgt cgtggcatcc gcaccatggc ttggaaaatc aggtgttccc
     4621 tggccgttag ggagccgata aagaggtcat cctgctggac ataggcgcac ctggcctgca
     4681 tctccttggc gtccacaggt tggccattga gcagtcgcat cccggatggc gatacttgga
     4741 tgccctgcgg cgatcgaaag gcaagggcat tcagcagggt cgtctttccg gcaccggaac
     4801 tgcccatcac ggccaaaagt tcgcccggat aggccacgcc gcaaactgag tttcaaattg
     4861 gtaattggac cctttattaa gatttcacac agatcagccg actgcgaata gaaactcacc
     4921 gttcttgagc aaatgtttcc tgggcgccgg tatgtgtcgc tcgttgcaga atagtccgcg
     4981 tgtccggttg accagctgcc gccatccgga gcccggctga ttgaccgccc caaagatgtc
     5041 catattgtgc caggcatagg tgaggttctc ggctagttgg ccgctccctg aaccggagtc
     5101 ctccggcgga ctgggtggcc ggagcgtgcc gtagtttttg gcctgcccga agccctggtt
     5161 aatgcagctc tgcgaagccg ctccgctgtc accctgcaat gataggggat ctcaaatatc
     5221 aactacaagc gttatgctca tctaaccccg aacaaaaagt accccgaagt atcctacgaa
     5281 gtaggtttat acttttattt attttttgtg catctaggat cagcttaaaa tatctggttg
     5341 ttatattttt tgtaaaaaag aatatagtcg aaaatgaatg cctttagatg tcttgatcat
     5401 gatatgatct caaaaattgt cttatatagc gagaacagct accagaataa tctgtttcgt
     5461 gtcactattt gtttgtgcaa ttgcggtttg ggatttttgt gggtcgcagt tctcacgccg
     5521 cagacaattt gatgttgcaa tcgcagttcc tatagatcaa gtgaacttaa gatgtatgca
     5581 catgtactac tcacattgtt cagatgctcg gcagatgggt gtttgctgcc tccgcgaatt
     5641 aatagctcct gatcctcttg gcccattgcc gggatttttc acactttccc ctgcttaccc
     5701 acccaaaacc aatcaccacc ccaatcactc aaaaaacaaa caaaaataag aagcgagagg
     5761 agttttggca cagcactttg tgtttaattg atggcgtaaa ccgcttggag cttcgtcacg
     5821 aaaccgctga caaaatgcaa ctgaaggcgg acattgacgc tacgtaacgc tacaaacggt
     5881 ggcgaaagag atagcggacg cagcggcgaa agagacggcg atatttctgt ggacagagaa
     5941 ggaggcaaac agcgctgact ttgagtggaa tgtcattttg agtgagaggt aatcgaaaga
     6001 acctggtaca tcaaataccc ttggatcgaa gtaaatttaa aactgatcag ataagttcaa
     6061 tgatatccag tgcagtaaaa aaaaaaaatg ttttttttat ctactttccg caaaaatggg
     6121 ttttattaac ttacatacat actaggggcc cggtacgtac ccaattcgcc ctatagtgag
     6181 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     6241 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     6301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     6361 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6421 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6481 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6541 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6601 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6661 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6721 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6781 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     6841 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6901 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6961 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     7021 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     7081 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     7141 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     7201 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     7261 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     7321 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     7381 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     7441 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7501 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7561 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7621 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7681 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7741 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7801 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7861 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7921 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7981 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     8041 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     8101 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     8161 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     8221 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     8281 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     8341 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     8401 acaggtatcc g
//
