LOCUS       sequence-12346-e        7963 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7963)
  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 7963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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
FEATURES             Location/Qualifiers
     source          1..7963
                     /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"
     protein_bind    complement(1383..1416)
                     /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(1601..1619)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(2071..2142,2544..2817,2892..3546,3608..3923,4127..4258,
                     4329..4943)
                     /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(5717..5750)
                     /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(5775..5795)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5775..5793)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5814..5830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5814..5830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5827..5849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5838..5854
                     /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(5862..5868,5869..5886,5887..5892))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5907..5928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6045..6062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6216..6804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6296..6315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6975..7766,7767..7835))
                     /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     7598..7617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7836..7940)
                     /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 gcacgcgtgg cgcgcctcta gatcgatacc gtcgactaaa gccaaataga aaattattca
     1141 gttcctggct taagttttta aaagtgatat tatttatttg gttgtaacca accaaaagaa
     1201 tgtaaataac taatacataa ttatgttagt tttaagttag caacaaattg attttagcta
     1261 tattagctac ttggttaata aatagaatat atttatttaa agataattgc gtttttattg
     1321 tcagggagtg agtttgctta aaaactcgtt tagatccact agcttaatta agcatgcact
     1381 agataacttc gtatagcata cattatacga agttatagct tctgatggaa ttagaacttg
     1441 gcaaaacaat actgagaatg aagtgtatgt ggaacagagg ctgctgatct cgttcttcag
     1501 gctatgaaac tgacacattt ggaaaccaca gtacttagaa ccacaaagtg ggaatcaaga
     1561 gaaaaacaat gatcccacga gagatcgaat tctagtatgt atgtaagtta ataaaaccca
     1621 tttttgcgga aagtagataa aaaaaacatt tttttttttt actgcactgg atatcattga
     1681 acttatctga tcagttttaa atttacttcg atccaagggt atttgatgta ccaggttctt
     1741 tcgattacct ctcactcaaa atgacattcc actcaaagtc agcgctgttt gcctccttct
     1801 ctgtccacag aaatatcgcc gtctctttcg ccgctgcgtc cgctatctct ttcgccaccg
     1861 tttgtagcgt tacgtagcgt caatgtccgc cttcagttgc attttgtcag cggtttcgtg
     1921 acgaagctcc aagcggttta cgccatcaat taaacacaaa gtgctgtgcc aaaactcctc
     1981 tcgcttctta tttttgtttg ttttttgagt gattggggtg gtgattggtt ttgggtgggt
     2041 aagcagggga aagtgtgaaa aatcccggca atgggccaag aggatcagga gctattaatt
     2101 cgcggaggca gcaaacaccc atctgccgag catctgaaca atgtgagtag tacatgtgca
     2161 tacatcttaa gttcacttga tctataggaa ctgcgattgc aacatcaaat tgtctgcggc
     2221 gtgagaactg cgacccacaa aaatcccaaa ccgcaattgc acaaacaaat agtgacacga
     2281 aacagattat tctggtagct gttctcgcta tataagacaa tttttgagat catatcatga
     2341 tcaagacatc taaaggcatt cattttcgac tatattcttt tttacaaaaa atataacaac
     2401 cagatatttt aagctgatcc tagatgcaca aaaaataaat aaaagtataa acctacttcg
     2461 taggatactt cggggtactt tttgttcggg gttagatgag cataacgctt gtagttgata
     2521 tttgagatcc cctatcattg cagggtgaca gcggagcggc ttcgcagagc tgcattaacc
     2581 agggcttcgg gcaggccaaa aactacggca cgctccggcc acccagtccg ccggaggact
     2641 ccggttcagg gagcggccaa ctagccgaga acctcaccta tgcctggcac aatatggaca
     2701 tctttggggc ggtcaatcag ccgggctccg gatggcggca gctggtcaac cggacacgcg
     2761 gactattctg caacgagcga cacataccgg cgcccaggaa acatttgctc aagaacggtg
     2821 agtttctatt cgcagtcggc tgatctgtgt gaaatcttaa taaagggtcc aattaccaat
     2881 ttgaaactca gtttgcggcg tggcctatcc gggcgaactt ttggccgtga tgggcagttc
     2941 cggtgccgga aagacgaccc tgctgaatgc ccttgccttt cgatcgccgc agggcatcca
     3001 agtatcgcca tccgggatgc gactgctcaa tggccaacct gtggacgcca aggagatgca
     3061 ggccaggtgc gcctatgtcc agcaggatga cctctttatc ggctccctaa cggccaggga
     3121 acacctgatt ttccaagcca tggtgcggat gccacgacat ctgacctatc ggcagcgagt
     3181 ggcccgcgtg gatcaggtga tccaggagct ttcgctcagc aaatgtcagc acacgatcat
     3241 cggtgtgccc ggcagggtga aaggtctgtc cggcggagaa aggaagcgtc tggcattcgc
     3301 ctccgaggct ctaaccgatc cgccgcttct gatctgcgat gagcccacct ccggactgga
     3361 ctcctttacc gcccacagcg tcgtccaggt gctgaagaag ctgtcgcaga agggcaagac
     3421 cgtcatcctg accattcatc agccgtcttc cgagctgttt gagctctttg acaagatcct
     3481 tctgatggcc gagggcaggg tagctttctt gggcactccc agcgaagccg tcgacttctt
     3541 ttcctagtga gttcgatgtg tttattaagg gtatctagta ttacataaca tctcaactcc
     3601 tatccagcgt gggtgcccag tgtcctacca actacaatcc ggcggacttt tacgtacagg
     3661 tgttggccgt tgtgcccgga cgggagatcg agtcccgtga tcggatcgcc aagatatgcg
     3721 acaattttgc cattagcaaa gtagcccggg atatggagca gttgttggcc accaaaaatc
     3781 tggagaagcc actggagcag ccggagaatg ggtacaccta caaggccacc tggttcatgc
     3841 agttccgggc ggtcctgtgg cgatcctggc tgtcggtgct caaggaacca ctcctcgtaa
     3901 aagtgcgact tattcagaca acggtgagtg gttccagtgg aaacaaatga tataacgctt
     3961 acaattcttg gaaacaaatt cgctagattt tagatagaat tgcctgattc cacacccttc
     4021 ttagtttttt tcaatgagat gtatagttta tagttttgca gaagataaat aaatttcatt
     4081 taactcgcga atattaatga gatgcgagta acattttaat ttgcagatgg ttgccatctt
     4141 gattggcctc atctttttgg gccaacaact cacgcaagtg ggtgtgatga atatcaacgg
     4201 agccatcttc ctcttcctga ccaacatgac ctttcaaaac gtctttgcca cgataaatgt
     4261 aagtcatgtt tagaatacat ttgcatttca ataatttact aactttctaa tgaatcgatt
     4321 cgatttaggt gttcacctca gagctgccag tttttatgag ggaggcccga agtcgacttt
     4381 atcgctgtga cacatacttt ctgggcaaaa cgattgccga attgccgctt tttctcacag
     4441 tgccactggt cttcacggcg attgcctatc cgatgatcgg actgcgggcc ggagtgctgc
     4501 acttcttcaa ctgcctggcg ctggtcactc tggtggccaa tgtgtcaacg tccttcggat
     4561 atctaatatc ctgcgccagc tcctcgacct cgatggcgct gtctgtgggt ccgccggtta
     4621 tcataccatt cctgctcttt ggcggcttct tcttgaactc gggctcggtg ccagtatacc
     4681 tcaaatggtt gtcgtacctc tcatggttcc gttacgccaa cgagggtctg ctgattaacc
     4741 aatgggcgga cgtggagccg ggcgaaatta gctgcacatc gtcgaacacc acgtgcccca
     4801 gttcgggcaa ggtcatcctg gagacgctta acttctccgc cgccgatctg ccgctggact
     4861 acgtgggtct ggccattctc atcgtgagct tccgggtgct cgcatatctg gctctaagac
     4921 ttcgggcccg acgcaaggag tagccgacat atatccgaaa taactgcttg tttttttttt
     4981 ttaccattat taccatcgtg tttactgttt attgccccct caaaaagcta atgtaattat
     5041 atttgtgcca ataaaaacaa gatatgacct atagaataca agtatttccc cttcgaacat
     5101 ccccacaagt agactttgga tttgtcttct aaccaaaaga cttacacacc tgcatacctt
     5161 acatcaaaaa ctcgtttatc gctacataaa acaccgggat atatttttta tatacatact
     5221 tttcaaatcg cgcgccctct tcataattca cctccaccac accacgtttc gtagttgctc
     5281 tttcgctgtc tcccacccgc tctccgcaac acattcacct tttgttcgac gaccttggag
     5341 cgactgtcgt tagttccgcg cgattcggtt cgctcaaatg gttccgagtg gttcatttcg
     5401 tctcaataga aattagtaat aaatatttgt atgtacaatt tatttgctcc aatatatttg
     5461 tatatatttc cctcacagct atatttattc taatttaata ttatgacttt ttaaggtaat
     5521 tttttgtgac ctgttcggag tgattagcgt tacaatttga actgaaagtg acatccagtg
     5581 tttgttcctt gtgtagatgc atctcaaaaa aatggtgggc ataatagtgt tgtttatata
     5641 tatcaaaaat aacaactata ataataagaa tacatttaat ttagaaaatg cttggatttc
     5701 actggaacta gaattcataa cttcgtatag catacattat acgaagttat ggatctgagc
     5761 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     5821 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5881 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5941 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6001 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6061 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7261 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7561 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7681 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
