LOCUS       sequence-47254-e        4546 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 4546)
  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 4546)
  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"; journalName: "PLoS 
            Genet."; date: "2011-10"; volume: "7(10)"; pages: "e1002308. Epub 
            2011 Oct 13"
COMMENT     Sequence Label: pPD95.79-wVenus
FEATURES             Location/Qualifiers
     source          1..4546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1986..2008)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2108..2127
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2205..2223)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2291..2395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2396..2464,2465..3256)
                     /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(2614..2633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3427..4015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3916..3935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4169..4186
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4303..4324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4339..4344,4345..4362,4363..4369)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4377..4393
                     /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     4382..4404
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4401..4417
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4401..4417
                     /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 cttgcttgga
      121 gggtaccggt agaaaaaatg tccaaaggtg aggagctttt cacaggagtt gtgccaattc
      181 tcgtcgaact cgatggtgac gttaatggac ataagttctc cgtatcggga gagggagaag
      241 gagatgctac ttatggaaag ttaaccctta agttgatctg tacaacggga aaacttccag
      301 tgccatgggt aagtttaaac atatatatac taactaaccc tgattattta aattttcagc
      361 caaccctggt tacaactctt ggatacggat tgcaatgctt cgcaagatac ccagatcaca
      421 tgaagcgaca tgacttcttc aaaagtgcca tgccagaggg atacgtgcag gaacgtacca
      481 ttttttttaa agacgatgga aattacaaaa caagtaagtt taaacagttc ggtactaact
      541 aaccatacat atttaaattt tcaggagccg aagtcaaatt tgagggagat actctcgtta
      601 atcgaatcga acttaagggt atcgacttca aagaagatgg aaacatcctt ggacacaagt
      661 tggaatacaa ttacaattcg cacaatgttt acattacagc agataagcaa aagaacggaa
      721 ttaaagcagt aagtttaaac atgattttac taactaacta atctgattta aattttcaga
      781 acttcaagat ccgtcacaat attgaggacg gaggagttca gttagctgat cattatcaac
      841 agaacactcc aatcggtgac ggtccagtac ttttgccaga caatcactac ctgtcctacc
      901 aatctgcact ctcgaaagat ccaaacgaaa agcgtgacca catggtcctt ttggagttcg
      961 tgaccgccgc tggaattaca ctgggaatgg atgaactcta caaatagcat tcgtagaatt
     1021 gtaagtttaa actttctcat actaactaac gcgctctatt taaattttca gaattccaac
     1081 tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt
     1141 catcagagta agtttaaact gagttctact aactaacgag taatatttaa attttcagca
     1201 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     1261 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     1321 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     1381 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     1441 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     1501 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     1561 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     1621 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     1681 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     1741 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     1801 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     1861 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     1921 ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     1981 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2041 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2101 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2161 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     2221 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2341 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2401 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2461 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2521 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2581 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2641 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2701 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2761 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2821 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2881 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2941 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3001 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3061 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3121 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3181 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3241 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3301 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3541 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3781 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     3841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4081 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4141 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4201 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4261 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4321 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4381 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctgtaagt
     4441 ttaaacatga tcttactaac taactattct catttaaatt ttcagagctt aaaaatggct
     4501 gaaatcactc acaacgatgg atacgctaac aacttggaaa tgaaat
//
