LOCUS       sequence-12375-e        8266 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 8266)
  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 8266)
  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: pBS130
FEATURES             Location/Qualifiers
     source          1..8266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..31)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15..31)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(28..50)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    39..55
                     /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(63..69,70..87,88..93))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(246..263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(417..1005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(497..516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1176..1967,1968..2036))
                     /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     1799..1818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2037..2141)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2209..2227
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2265..2287)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2387..2406
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2600..2622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2614..2631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2615..2631
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2870..2889
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             4183..5997
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     CDS             5998..6018
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
       61 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      121 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      181 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
      241 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      301 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      361 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      421 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      481 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      541 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      601 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      661 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      721 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      781 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      841 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      901 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      961 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1021 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1081 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1141 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1201 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1261 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1321 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1381 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1441 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1501 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1561 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1621 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1681 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1741 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1801 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1861 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1921 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1981 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2041 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2101 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2161 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2221 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2281 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2341 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     2401 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2461 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2521 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2581 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
     2641 gcatgcctgc aggtcgactc tagaggatcc gtcgacgctc tcccttatgc gactcctgca
     2701 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     2761 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     2821 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     2881 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     2941 cgtagaggat cttgaatatg acctacaaaa acgtaaccat tccataaaaa cagtctccaa
     3001 ctcgtagaca caagtcaatg aggaacatac tacgtactaa aacttttaaa attttaatta
     3061 aataatttaa aattttagga ttaaacttcg atttagtgtt ttgtgtaaag agtttcttga
     3121 cagtctcaaa agtgtgtaaa cttggaattt gtggtttatt ttcttgtata attaatattt
     3181 ttaatttatt ttttgattta taagtaaaat aaatattgtt taattatatt ttataaaaaa
     3241 attgcgttta attaagcaaa aaacccttaa tttttagctt taaagtcaaa aattcaactt
     3301 atttcacagt gtgtaaacag tttcttgaca aactgtatat ctaatatttt taattttttt
     3361 tttatttata agtaaaataa atattgttta attatattaa aaaaaaaaat tccgtttaat
     3421 taagcaaata acccttaatt tttaccttta aagtcaaaaa ttcgacatat ttcacagttt
     3481 gtaaacttga taaactgtat gtaatagtct tggctttcct catatgtaca tatgtatgta
     3541 aatatgtaaa ataagtcgca actaaattct aatacatttt tcagagtctt aaattaattt
     3601 tatcgtatat taaaacagaa gaaagtccgt taatagttga tttcattaac taaaagtaca
     3661 aaataatctt taatacatat gccgatcaga catttattgg tttagaagcg cagtattttt
     3721 tttggcggaa tacgcataac aaagcgcttc gattatcttt aacataagtt atttaagcag
     3781 ccgtatttat aaagaaattt ccaaaataaa gcgaatattc tagaatccca aaacaaactg
     3841 gttattgtgg taggtcattt gtttggcaga aagaaaactc gagaaatttc tctggccgtt
     3901 attcgttatt ctctcttttc tttttgggtc tctccctctc tgcactaatg ctctctcact
     3961 ctgtcacaca gtaaacggca tactgctctc gttggttcga gagagcgcgc ctcgaatgtt
     4021 cgcgaaaaga gcgccggagt ataaatagag gcgcttcgtc tacggagcga caattcaatt
     4081 caaacaagca aagtgaacac gtcgctaagc gaaagctaag caaataaaca agcgcagctg
     4141 aacaagctaa acaatctgca ggtcgactct agaggatcca aaatggacac gtacgcgggt
     4201 gcttacgacc gtcagtcgcg cgagcgcgaa aattcgagcg cagcaagccc agcgacacag
     4261 cgtagcgcca acgaagacaa ggcggccgac cttcagcgcg aagtcgagcg cgacgggggc
     4321 cggttcaggt tcgtcgggca tttcagcgaa gcgccgggca cgtcggcgtt cgggacggcg
     4381 gagcgcccgg agttcgaacg catcctgaac gaatgccgcg ccgggcggct caacatgatc
     4441 attgtctatg acgtgtcgcg cttctcgcgc ctgaaggtca tggacgcgat tccgattgtc
     4501 tcggaattgc tcgccctggg cgtgacgatt gtttccactc aggaaggcgt cttccggcag
     4561 ggaaacgtca tggacctgat tcacctgatt atgcggctcg acgcgtcgca caaagaatct
     4621 tcgctgaagt cggcgaagat tctcgacacg aagaaccttc agcgcgaatt gggcgggtac
     4681 gtcggcggga aggcgcctta cggcttcgag cttgtttcgg agacgaagga gatcacgcgc
     4741 aacggccgaa tggtcaatgt cgtcatcaac aagcttgcgc actcgaccac tccccttacc
     4801 ggacccttcg agttcgagcc cgacgtaatc cggtggtggt ggcgtgagat caagacgcac
     4861 aaacaccttc ccttcaagcc gggcagtcaa gccgccattc acccgggcag catcacgggg
     4921 ctttgtaagc gcatggacgc tgacgccgtg ccgacccggg gcgagacgat tgggaagaag
     4981 accgcttcaa gcgcctggga cccggcaacc gttatgcgaa tccttcggga cccgcgtatt
     5041 gcgggcttcg ccgctgaggt gatctacaag aagaagccgg acggcacgcc gaccacgaag
     5101 attgagggtt accgcattca gcgcgacccg atcacgctcc ggccggtcga gcttgattgc
     5161 ggaccgatca tcgagcccgc tgagtggtat gagcttcagg cgtggttgga cggcaggggg
     5221 cgcggcaagg ggctttcccg ggggcaagcc attctgtccg ccatggacaa gctgtactgc
     5281 gagtgtggcg ccgtcatgac ttcgaagcgc ggggaagaat cgatcaagga ctcttaccgc
     5341 tgccgtcgcc ggaaggtggt cgacccgtcc gcacctgggc agcacgaagg cacgtgcaac
     5401 gtcagcatgg cggcactcga caagttcgtt gcggaacgca tcttcaacaa gatcaggcac
     5461 gccgaaggcg acgaagagac gttggcgctt ctgtgggaag ccgcccgacg cttcggcaag
     5521 ctcactgagg cgcctgagaa gagcggcgaa cgggcgaacc ttgttgcgga gcgcgccgac
     5581 gccctgaacg cccttgaaga gctgtacgaa gaccgcgcgg caggcgcgta cgacggaccc
     5641 gttggcagga agcacttccg gaagcaacag gcagcgctga cgctccggca gcaaggggcg
     5701 gaagagcggc ttgccgaact tgaagccgcc gaagccccga agcttcccct tgaccaatgg
     5761 ttccccgaag acgccgacgc tgacccgacc ggccctaagt cgtggtgggg gcgcgcgtca
     5821 gtagacgaca agcgcgtgtt cgtcgggctc ttcgtagaca agatcgttgt cacgaagtcg
     5881 actacgggca gggggcaggg aacgcccatc gagaagcgcg cttcgatcac gtgggcgaag
     5941 ccgccgaccg acgacgacga agacgacgcc caggacggca cggaagacgt agcggcgcct
     6001 aagaagaaga ggaaggttta gctcgaattc tggataaacc caaaaggctc ccatttcaaa
     6061 aaggtatcat tgtgaataat gcttcgcttg atggcttgta taaatatttg caagaaaact
     6121 tcagtatgca atacatatta acaagccgtc tcaaccaaga cattgtggag catttttttg
     6181 gcagcatgcg atcgagaggt ggacaattcg accatcccac tccactgcag tttaagtata
     6241 ggttaagaaa atatataata gccaggaata cagaaatgtt aagaaattcg ggaaatatcg
     6301 aagaggacaa ctctgaaagc tggcttaatt tagatttcag ttctaaagaa aacgaaaata
     6361 aaagtaaaga tgatgagcct gtcgatgatg agcctgtcga tgagatgtta agcaatatag
     6421 atttcaccga aatggatgag ttgacggagg atgcgatgga atatatcgcg ggctatgtca
     6481 ttaaaaaatt gagaatcagt gacaaagtaa aagaaaattt gacatttaca tacgtcgacg
     6541 aggtgtctca cggcggactt attaagccgt ccgaaaaatt tcaagagaag ttaaaagagc
     6601 tagaatgtat ttttttgcat tatacaaata ataataattt tgaaattaca aataatgtaa
     6661 aggaaaaatt aatattagca gcgcgaaacg tcgatgttga taaacaagta aaatcttttt
     6721 attttaaaat tagaaatata ttttagaatt aagtacttca acaaaaaaaa ttgaaattaa
     6781 aaatcaaaaa caaaagttaa ttggaaactc caaattatta aaaataaaac tttaaaaata
     6841 atttcgtcta attaatatta tgagttaatt caaaccccac ggacatgcta agggttaatc
     6901 aacaatcata tcgctgtctc actcagactc aatacgacac tcagaatact attcctttca
     6961 ctcgcactta ttgcaagcat acgttaagtg gatgtctctt gccgacggga cggtcgatcg
     7021 acctgcagcc aagctagctt cttttgattg agcttatatt ttcgggtagc agtcgcgccg
     7081 aaatcggaac tcatatttca attcatcgga aaacttgctc aactttcatt aatatactca
     7141 aaataaatag tgactaagac ttttcaattg aaaatcagaa tgcattgcat gccacgagat
     7201 tctattgaat aatagataga acaaaaaagt aaccattgaa tttttcttaa tctaaaagga
     7261 atatttaata ttaatcagtg gtatataaat ttgttcaaac agctctccag cttagtttac
     7321 acattaaacg actcaattat ttttaaaaca tttaattcac tatgtatata taaacaaatt
     7381 aacaaataaa atcgtcattt taaatgctta agcagtacaa acaataaaaa caattttaag
     7441 gcacaatgtt ccatggcgta aatgatcctg gttcgggtat ttcaagctat aattaataaa
     7501 ttaataataa aagttagttt aatgttctaa actaatccaa gggtaactta ccagttccgt
     7561 gaacttatcc tgacaagcaa ttcgaatgcg tgccgatgtg gaaggcgcct ccagtgggaa
     7621 gtcaccactc aagccctgat cctcgcgagc agctgcaatg gcctccttaa tggcaaagaa
     7681 tgcagatgat ccaatgaaga gcggaggttc acccactgcc ttggaagagt agactgcccg
     7741 tggattgggg gcaccggtca gtaggctgac attgaactcc ccgggaatgt cggcaaatcc
     7801 tggcagcttg tacatgcccg gacctctgga gtaaagcatg ccttgtggtg agtacatgag
     7861 ttcctccaaa gtgaacagtc catagccctg catgaatgct ccctcgatct gaccaatgtc
     7921 aatagccgga ttcaggctag agccgatgtc catcacgatg tctgtgctga gcacctgatg
     7981 gtcgccagtc aggcaatcga tctctaccac agtgactccc acgccattcg tgtagtagct
     8041 ataggtgcga gcattgggat tcgtttccgg gtgatatcca atcccgggca tggcatagaa
     8101 tcctgtggcc gagaggctga cccgatcgaa atacgccttg ttgatccact ccttccaggt
     8161 gcctccaggc aatgcctcct tgatgggcgc cagtcttttg ttcaactttt cacacgcatc
     8221 cagtacggcc attccgttca gatcggatcc ccgggtaccg agctcg
//
