LOCUS       sequence-47253-e        4541 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 4541)
  AUTHORS   Yamamoto Y, Takeshita H, Sawa H
  TITLE     Multiple Wnts redundantly control polarity orientation in 
            Caenorhabditis elegans epithelial stem cells.
  JOURNAL   PLoS Genet. 2011 Oct;7(10):e1002308. Epub 2011 Oct 13.
  PUBMED    22022276
REFERENCE   2  (bases 1 to 4541)
  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: "PLoS 
            Genet."; date: "2011-10"; volume: "7(10)"; pages: "e1002308. Epub 
            2011 Oct 13"
COMMENT     Sequence Label: pPD95.77-wVenus
FEATURES             Location/Qualifiers
     source          1..4541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1981..2003)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2103..2122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2200..2218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2286..2390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2391..2459,2460..3251)
                     /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(2609..2628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3422..4010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3911..3930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4164..4181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4298..4319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4334..4339,4340..4357,4358..4364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4372..4388
                     /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     4377..4399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4396..4412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4396..4412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttgagggta
      121 ccggtagaaa aaatgtccaa aggtgaggag cttttcacag gagttgtgcc aattctcgtc
      181 gaactcgatg gtgacgttaa tggacataag ttctccgtat cgggagaggg agaaggagat
      241 gctacttatg gaaagttaac ccttaagttg atctgtacaa cgggaaaact tccagtgcca
      301 tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt tcagccaacc
      361 ctggttacaa ctcttggata cggattgcaa tgcttcgcaa gatacccaga tcacatgaag
      421 cgacatgact tcttcaaaag tgccatgcca gagggatacg tgcaggaacg taccattttt
      481 tttaaagacg atggaaatta caaaacaagt aagtttaaac agttcggtac taactaacca
      541 tacatattta aattttcagg agccgaagtc aaatttgagg gagatactct cgttaatcga
      601 atcgaactta agggtatcga cttcaaagaa gatggaaaca tccttggaca caagttggaa
      661 tacaattaca attcgcacaa tgtttacatt acagcagata agcaaaagaa cggaattaaa
      721 gcagtaagtt taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc
      781 aagatccgtc acaatattga ggacggagga gttcagttag ctgatcatta tcaacagaac
      841 actccaatcg gtgacggtcc agtacttttg ccagacaatc actacctgtc ctaccaatct
      901 gcactctcga aagatccaaa cgaaaagcgt gaccacatgg tccttttgga gttcgtgacc
      961 gccgctggaa ttacactggg aatggatgaa ctctacaaat agcattcgta gaattgtaag
     1021 tttaaacttt ctcatactaa ctaacgcgct ctatttaaat tttcagaatt ccaactgagc
     1081 gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca
     1141 gagtaagttt aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg
     1201 cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc
     1261 cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt
     1321 tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa
     1381 ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc
     1441 caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt
     1501 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     1561 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     1621 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     1681 attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc
     1741 gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg
     1801 tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg
     1861 tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt
     1921 tcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     2041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     2101 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     2161 atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt
     2221 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2941 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3001 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3241 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3301 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3361 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3421 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3481 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3541 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3781 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     3841 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3901 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3961 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4021 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4081 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4141 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4201 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4261 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4321 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4381 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa
     4441 catgatctta ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat
     4501 cactcacaac gatggatacg ctaacaactt ggaaatgaaa t
//
