LOCUS       sequence-12351-e        8445 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 8445)
  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 8445)
  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-VP16AD
FEATURES             Location/Qualifiers
     source          1..8445
                     /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             1118..1138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1151..1384
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     protein_bind    complement(1865..1898)
                     /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(2083..2101)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(2553..2624,3026..3299,3374..4028,4090..4405,4609..4740,
                     4811..5425)
                     /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(6199..6232)
                     /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(6257..6277)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6257..6275)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6296..6312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6296..6312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6309..6331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6320..6336
                     /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(6344..6350,6351..6368,6369..6374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6389..6410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6527..6544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6698..7286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6778..6797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7457..8248,8249..8317))
                     /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     8080..8099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8318..8422)
                     /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 gataaagcgg aattaattcc cgagcctcca aaaaagaaga gaaaggtcga
     1141 attgggtacc gcccccccga ccgatgtcag cctgggggac gagctccact tagacggcga
     1201 ggacgtggcg atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga
     1261 cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga
     1321 tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg
     1381 tggggctagc ggaggaggtg gtggaggtgg aggtggaggt actagtctgg agatcgaggc
     1441 cgccttcctg gagcgcgaga acaccgccct ggagacccgc gtggccgagc tgcgccagcg
     1501 cgtgcagcgc ctgcgcaacc gcgtgagcca gaaccgcacc cgctacggcc ccctgggcgg
     1561 cggcaagtaa ggcgcgcctc tagatcgata ccgtcgacta aagccaaata gaaaattatt
     1621 cagttcctgg cttaagtttt taaaagtgat attatttatt tggttgtaac caaccaaaag
     1681 aatgtaaata actaatacat aattatgtta gttttaagtt agcaacaaat tgattttagc
     1741 tatattagct acttggttaa taaatagaat atatttattt aaagataatt gcgtttttat
     1801 tgtcagggag tgagtttgct taaaaactcg tttagatcca ctagcttaat taagcatgca
     1861 ctagataact tcgtatagca tacattatac gaagttatag cttctgatgg aattagaact
     1921 tggcaaaaca atactgagaa tgaagtgtat gtggaacaga ggctgctgat ctcgttcttc
     1981 aggctatgaa actgacacat ttggaaacca cagtacttag aaccacaaag tgggaatcaa
     2041 gagaaaaaca atgatcccac gagagatcga attctagtat gtatgtaagt taataaaacc
     2101 catttttgcg gaaagtagat aaaaaaaaca tttttttttt ttactgcact ggatatcatt
     2161 gaacttatct gatcagtttt aaatttactt cgatccaagg gtatttgatg taccaggttc
     2221 tttcgattac ctctcactca aaatgacatt ccactcaaag tcagcgctgt ttgcctcctt
     2281 ctctgtccac agaaatatcg ccgtctcttt cgccgctgcg tccgctatct ctttcgccac
     2341 cgtttgtagc gttacgtagc gtcaatgtcc gccttcagtt gcattttgtc agcggtttcg
     2401 tgacgaagct ccaagcggtt tacgccatca attaaacaca aagtgctgtg ccaaaactcc
     2461 tctcgcttct tatttttgtt tgttttttga gtgattgggg tggtgattgg ttttgggtgg
     2521 gtaagcaggg gaaagtgtga aaaatcccgg caatgggcca agaggatcag gagctattaa
     2581 ttcgcggagg cagcaaacac ccatctgccg agcatctgaa caatgtgagt agtacatgtg
     2641 catacatctt aagttcactt gatctatagg aactgcgatt gcaacatcaa attgtctgcg
     2701 gcgtgagaac tgcgacccac aaaaatccca aaccgcaatt gcacaaacaa atagtgacac
     2761 gaaacagatt attctggtag ctgttctcgc tatataagac aatttttgag atcatatcat
     2821 gatcaagaca tctaaaggca ttcattttcg actatattct tttttacaaa aaatataaca
     2881 accagatatt ttaagctgat cctagatgca caaaaaataa ataaaagtat aaacctactt
     2941 cgtaggatac ttcggggtac tttttgttcg gggttagatg agcataacgc ttgtagttga
     3001 tatttgagat cccctatcat tgcagggtga cagcggagcg gcttcgcaga gctgcattaa
     3061 ccagggcttc gggcaggcca aaaactacgg cacgctccgg ccacccagtc cgccggagga
     3121 ctccggttca gggagcggcc aactagccga gaacctcacc tatgcctggc acaatatgga
     3181 catctttggg gcggtcaatc agccgggctc cggatggcgg cagctggtca accggacacg
     3241 cggactattc tgcaacgagc gacacatacc ggcgcccagg aaacatttgc tcaagaacgg
     3301 tgagtttcta ttcgcagtcg gctgatctgt gtgaaatctt aataaagggt ccaattacca
     3361 atttgaaact cagtttgcgg cgtggcctat ccgggcgaac ttttggccgt gatgggcagt
     3421 tccggtgccg gaaagacgac cctgctgaat gcccttgcct ttcgatcgcc gcagggcatc
     3481 caagtatcgc catccgggat gcgactgctc aatggccaac ctgtggacgc caaggagatg
     3541 caggccaggt gcgcctatgt ccagcaggat gacctcttta tcggctccct aacggccagg
     3601 gaacacctga ttttccaagc catggtgcgg atgccacgac atctgaccta tcggcagcga
     3661 gtggcccgcg tggatcaggt gatccaggag ctttcgctca gcaaatgtca gcacacgatc
     3721 atcggtgtgc ccggcagggt gaaaggtctg tccggcggag aaaggaagcg tctggcattc
     3781 gcctccgagg ctctaaccga tccgccgctt ctgatctgcg atgagcccac ctccggactg
     3841 gactccttta ccgcccacag cgtcgtccag gtgctgaaga agctgtcgca gaagggcaag
     3901 accgtcatcc tgaccattca tcagccgtct tccgagctgt ttgagctctt tgacaagatc
     3961 cttctgatgg ccgagggcag ggtagctttc ttgggcactc ccagcgaagc cgtcgacttc
     4021 ttttcctagt gagttcgatg tgtttattaa gggtatctag tattacataa catctcaact
     4081 cctatccagc gtgggtgccc agtgtcctac caactacaat ccggcggact tttacgtaca
     4141 ggtgttggcc gttgtgcccg gacgggagat cgagtcccgt gatcggatcg ccaagatatg
     4201 cgacaatttt gccattagca aagtagcccg ggatatggag cagttgttgg ccaccaaaaa
     4261 tctggagaag ccactggagc agccggagaa tgggtacacc tacaaggcca cctggttcat
     4321 gcagttccgg gcggtcctgt ggcgatcctg gctgtcggtg ctcaaggaac cactcctcgt
     4381 aaaagtgcga cttattcaga caacggtgag tggttccagt ggaaacaaat gatataacgc
     4441 ttacaattct tggaaacaaa ttcgctagat tttagataga attgcctgat tccacaccct
     4501 tcttagtttt tttcaatgag atgtatagtt tatagttttg cagaagataa ataaatttca
     4561 tttaactcgc gaatattaat gagatgcgag taacatttta atttgcagat ggttgccatc
     4621 ttgattggcc tcatcttttt gggccaacaa ctcacgcaag tgggtgtgat gaatatcaac
     4681 ggagccatct tcctcttcct gaccaacatg acctttcaaa acgtctttgc cacgataaat
     4741 gtaagtcatg tttagaatac atttgcattt caataattta ctaactttct aatgaatcga
     4801 ttcgatttag gtgttcacct cagagctgcc agtttttatg agggaggccc gaagtcgact
     4861 ttatcgctgt gacacatact ttctgggcaa aacgattgcc gaattgccgc tttttctcac
     4921 agtgccactg gtcttcacgg cgattgccta tccgatgatc ggactgcggg ccggagtgct
     4981 gcacttcttc aactgcctgg cgctggtcac tctggtggcc aatgtgtcaa cgtccttcgg
     5041 atatctaata tcctgcgcca gctcctcgac ctcgatggcg ctgtctgtgg gtccgccggt
     5101 tatcatacca ttcctgctct ttggcggctt cttcttgaac tcgggctcgg tgccagtata
     5161 cctcaaatgg ttgtcgtacc tctcatggtt ccgttacgcc aacgagggtc tgctgattaa
     5221 ccaatgggcg gacgtggagc cgggcgaaat tagctgcaca tcgtcgaaca ccacgtgccc
     5281 cagttcgggc aaggtcatcc tggagacgct taacttctcc gccgccgatc tgccgctgga
     5341 ctacgtgggt ctggccattc tcatcgtgag cttccgggtg ctcgcatatc tggctctaag
     5401 acttcgggcc cgacgcaagg agtagccgac atatatccga aataactgct tgtttttttt
     5461 ttttaccatt attaccatcg tgtttactgt ttattgcccc ctcaaaaagc taatgtaatt
     5521 atatttgtgc caataaaaac aagatatgac ctatagaata caagtatttc cccttcgaac
     5581 atccccacaa gtagactttg gatttgtctt ctaaccaaaa gacttacaca cctgcatacc
     5641 ttacatcaaa aactcgttta tcgctacata aaacaccggg atatattttt tatatacata
     5701 cttttcaaat cgcgcgccct cttcataatt cacctccacc acaccacgtt tcgtagttgc
     5761 tctttcgctg tctcccaccc gctctccgca acacattcac cttttgttcg acgaccttgg
     5821 agcgactgtc gttagttccg cgcgattcgg ttcgctcaaa tggttccgag tggttcattt
     5881 cgtctcaata gaaattagta ataaatattt gtatgtacaa tttatttgct ccaatatatt
     5941 tgtatatatt tccctcacag ctatatttat tctaatttaa tattatgact ttttaaggta
     6001 attttttgtg acctgttcgg agtgattagc gttacaattt gaactgaaag tgacatccag
     6061 tgtttgttcc ttgtgtagat gcatctcaaa aaaatggtgg gcataatagt gttgtttata
     6121 tatatcaaaa ataacaacta taataataag aatacattta atttagaaaa tgcttggatt
     6181 tcactggaac tagaattcat aacttcgtat agcatacatt atacgaagtt atggatctga
     6241 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     6301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
