LOCUS       sequence-12348-e        8592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8592)
  AUTHORS   Gohl DM, Silies MA, Gao XJ, Bhalerao S, Luongo FJ, Lin CC, Potter 
            CJ, Clandinin TR
  TITLE     A versatile in vivo system for directed dissection of gene 
            expression patterns.
  JOURNAL   Nat Methods. 2011 Mar;8(3):231-7. doi: 10.1038/nmeth.1561.
  PUBMED    21473015
REFERENCE   2  (bases 1 to 8592)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-03"; volume: "8"; issue: 
            "3"; pages: "231-7"
COMMENT     Sequence Label: pBPHLWL-GAL4DBD
FEATURES             Location/Qualifiers
     source          1..8592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    689..758
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     949..965
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1274..1714
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    complement(2012..2045)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2230..2248)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(2700..2771,3173..3446,3521..4175,4237..4552,4756..4887,
                     4958..5572)
                     /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    complement(6346..6379)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6404..6424)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6404..6422)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6443..6459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6443..6459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6456..6478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6467..6483
                     /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(6491..6497,6498..6515,6516..6521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6536..6557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6674..6691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6845..7433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6925..6944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7604..8395,8396..8464))
                     /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     8227..8246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8465..8569)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gacgatgtag gtcacggtct cgaagccgcg gtgcgggtgc cagggcgtgc
      721 ccttgggctc cccgggcgcg tactccacct cacccatctg gtccatcatg atgaacgggt
      781 cgaggtggcg gtagttgatc ccggcgaacg cgcggcgcac cgggaagccc tcgccctcga
      841 aaccgctggg cgcggtggtc acggtgagca cgggacgtgc gacggcgtcg gcgggtgcgg
      901 atacgcgggg cagcgtcagc gggttctcga cggtcacggc gggcatgtcg aggtcgacgg
      961 tatcgataag cttaccgaag tatacactta aattcagtgc acgtttgctt gttgagagga
     1021 aaggttgtgt gcggacgaat ttttttttga aaacattaac ccttacgtgg atccgcggcc
     1081 gcaaaacatg ctggagatcc gcgccgcctt cctgcgccag cgcaacaccg ccctgcgcac
     1141 cgaggtggcc gagctggagc aggaggtgca gcgcctggag aacgaggtga gccagtacga
     1201 gacccgctac ggccccctgg gcggcggcaa ggctagcgga ggaggtggtg gaggtggagg
     1261 tggaggtact agtatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
     1321 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
     1381 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
     1441 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
     1501 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
     1561 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
     1621 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
     1681 tagtaacaaa ggtcaaagac agttgactgt atcgtaaggc gcgcctctag atcgataccg
     1741 tcgactaaag ccaaatagaa aattattcag ttcctggctt aagtttttaa aagtgatatt
     1801 atttatttgg ttgtaaccaa ccaaaagaat gtaaataact aatacataat tatgttagtt
     1861 ttaagttagc aacaaattga ttttagctat attagctact tggttaataa atagaatata
     1921 tttatttaaa gataattgcg tttttattgt cagggagtga gtttgcttaa aaactcgttt
     1981 agatccacta gcttaattaa gcatgcacta gataacttcg tatagcatac attatacgaa
     2041 gttatagctt ctgatggaat tagaacttgg caaaacaata ctgagaatga agtgtatgtg
     2101 gaacagaggc tgctgatctc gttcttcagg ctatgaaact gacacatttg gaaaccacag
     2161 tacttagaac cacaaagtgg gaatcaagag aaaaacaatg atcccacgag agatcgaatt
     2221 ctagtatgta tgtaagttaa taaaacccat ttttgcggaa agtagataaa aaaaacattt
     2281 ttttttttta ctgcactgga tatcattgaa cttatctgat cagttttaaa tttacttcga
     2341 tccaagggta tttgatgtac caggttcttt cgattacctc tcactcaaaa tgacattcca
     2401 ctcaaagtca gcgctgtttg cctccttctc tgtccacaga aatatcgccg tctctttcgc
     2461 cgctgcgtcc gctatctctt tcgccaccgt ttgtagcgtt acgtagcgtc aatgtccgcc
     2521 ttcagttgca ttttgtcagc ggtttcgtga cgaagctcca agcggtttac gccatcaatt
     2581 aaacacaaag tgctgtgcca aaactcctct cgcttcttat ttttgtttgt tttttgagtg
     2641 attggggtgg tgattggttt tgggtgggta agcaggggaa agtgtgaaaa atcccggcaa
     2701 tgggccaaga ggatcaggag ctattaattc gcggaggcag caaacaccca tctgccgagc
     2761 atctgaacaa tgtgagtagt acatgtgcat acatcttaag ttcacttgat ctataggaac
     2821 tgcgattgca acatcaaatt gtctgcggcg tgagaactgc gacccacaaa aatcccaaac
     2881 cgcaattgca caaacaaata gtgacacgaa acagattatt ctggtagctg ttctcgctat
     2941 ataagacaat ttttgagatc atatcatgat caagacatct aaaggcattc attttcgact
     3001 atattctttt ttacaaaaaa tataacaacc agatatttta agctgatcct agatgcacaa
     3061 aaaataaata aaagtataaa cctacttcgt aggatacttc ggggtacttt ttgttcgggg
     3121 ttagatgagc ataacgcttg tagttgatat ttgagatccc ctatcattgc agggtgacag
     3181 cggagcggct tcgcagagct gcattaacca gggcttcggg caggccaaaa actacggcac
     3241 gctccggcca cccagtccgc cggaggactc cggttcaggg agcggccaac tagccgagaa
     3301 cctcacctat gcctggcaca atatggacat ctttggggcg gtcaatcagc cgggctccgg
     3361 atggcggcag ctggtcaacc ggacacgcgg actattctgc aacgagcgac acataccggc
     3421 gcccaggaaa catttgctca agaacggtga gtttctattc gcagtcggct gatctgtgtg
     3481 aaatcttaat aaagggtcca attaccaatt tgaaactcag tttgcggcgt ggcctatccg
     3541 ggcgaacttt tggccgtgat gggcagttcc ggtgccggaa agacgaccct gctgaatgcc
     3601 cttgcctttc gatcgccgca gggcatccaa gtatcgccat ccgggatgcg actgctcaat
     3661 ggccaacctg tggacgccaa ggagatgcag gccaggtgcg cctatgtcca gcaggatgac
     3721 ctctttatcg gctccctaac ggccagggaa cacctgattt tccaagccat ggtgcggatg
     3781 ccacgacatc tgacctatcg gcagcgagtg gcccgcgtgg atcaggtgat ccaggagctt
     3841 tcgctcagca aatgtcagca cacgatcatc ggtgtgcccg gcagggtgaa aggtctgtcc
     3901 ggcggagaaa ggaagcgtct ggcattcgcc tccgaggctc taaccgatcc gccgcttctg
     3961 atctgcgatg agcccacctc cggactggac tcctttaccg cccacagcgt cgtccaggtg
     4021 ctgaagaagc tgtcgcagaa gggcaagacc gtcatcctga ccattcatca gccgtcttcc
     4081 gagctgtttg agctctttga caagatcctt ctgatggccg agggcagggt agctttcttg
     4141 ggcactccca gcgaagccgt cgacttcttt tcctagtgag ttcgatgtgt ttattaaggg
     4201 tatctagtat tacataacat ctcaactcct atccagcgtg ggtgcccagt gtcctaccaa
     4261 ctacaatccg gcggactttt acgtacaggt gttggccgtt gtgcccggac gggagatcga
     4321 gtcccgtgat cggatcgcca agatatgcga caattttgcc attagcaaag tagcccggga
     4381 tatggagcag ttgttggcca ccaaaaatct ggagaagcca ctggagcagc cggagaatgg
     4441 gtacacctac aaggccacct ggttcatgca gttccgggcg gtcctgtggc gatcctggct
     4501 gtcggtgctc aaggaaccac tcctcgtaaa agtgcgactt attcagacaa cggtgagtgg
     4561 ttccagtgga aacaaatgat ataacgctta caattcttgg aaacaaattc gctagatttt
     4621 agatagaatt gcctgattcc acacccttct tagttttttt caatgagatg tatagtttat
     4681 agttttgcag aagataaata aatttcattt aactcgcgaa tattaatgag atgcgagtaa
     4741 cattttaatt tgcagatggt tgccatcttg attggcctca tctttttggg ccaacaactc
     4801 acgcaagtgg gtgtgatgaa tatcaacgga gccatcttcc tcttcctgac caacatgacc
     4861 tttcaaaacg tctttgccac gataaatgta agtcatgttt agaatacatt tgcatttcaa
     4921 taatttacta actttctaat gaatcgattc gatttaggtg ttcacctcag agctgccagt
     4981 ttttatgagg gaggcccgaa gtcgacttta tcgctgtgac acatactttc tgggcaaaac
     5041 gattgccgaa ttgccgcttt ttctcacagt gccactggtc ttcacggcga ttgcctatcc
     5101 gatgatcgga ctgcgggccg gagtgctgca cttcttcaac tgcctggcgc tggtcactct
     5161 ggtggccaat gtgtcaacgt ccttcggata tctaatatcc tgcgccagct cctcgacctc
     5221 gatggcgctg tctgtgggtc cgccggttat cataccattc ctgctctttg gcggcttctt
     5281 cttgaactcg ggctcggtgc cagtatacct caaatggttg tcgtacctct catggttccg
     5341 ttacgccaac gagggtctgc tgattaacca atgggcggac gtggagccgg gcgaaattag
     5401 ctgcacatcg tcgaacacca cgtgccccag ttcgggcaag gtcatcctgg agacgcttaa
     5461 cttctccgcc gccgatctgc cgctggacta cgtgggtctg gccattctca tcgtgagctt
     5521 ccgggtgctc gcatatctgg ctctaagact tcgggcccga cgcaaggagt agccgacata
     5581 tatccgaaat aactgcttgt tttttttttt taccattatt accatcgtgt ttactgttta
     5641 ttgccccctc aaaaagctaa tgtaattata tttgtgccaa taaaaacaag atatgaccta
     5701 tagaatacaa gtatttcccc ttcgaacatc cccacaagta gactttggat ttgtcttcta
     5761 accaaaagac ttacacacct gcatacctta catcaaaaac tcgtttatcg ctacataaaa
     5821 caccgggata tattttttat atacatactt ttcaaatcgc gcgccctctt cataattcac
     5881 ctccaccaca ccacgtttcg tagttgctct ttcgctgtct cccacccgct ctccgcaaca
     5941 cattcacctt ttgttcgacg accttggagc gactgtcgtt agttccgcgc gattcggttc
     6001 gctcaaatgg ttccgagtgg ttcatttcgt ctcaatagaa attagtaata aatatttgta
     6061 tgtacaattt atttgctcca atatatttgt atatatttcc ctcacagcta tatttattct
     6121 aatttaatat tatgactttt taaggtaatt ttttgtgacc tgttcggagt gattagcgtt
     6181 acaatttgaa ctgaaagtga catccagtgt ttgttccttg tgtagatgca tctcaaaaaa
     6241 atggtgggca taatagtgtt gtttatatat atcaaaaata acaactataa taataagaat
     6301 acatttaatt tagaaaatgc ttggatttca ctggaactag aattcataac ttcgtatagc
     6361 atacattata cgaagttatg gatctgagct ccagcttttg ttccctttag tgagggttaa
     6421 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6481 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6541 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6601 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6661 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6721 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6781 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7081 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7261 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7501 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7561 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7621 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7681 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7741 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7801 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7861 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7921 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7981 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8041 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8101 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8161 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8221 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8281 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8341 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8401 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8461 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8521 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8581 gaaaagtgcc ac
//
